Light measurement apparatus for reaction vessel and light measurement method

ABSTRACT

The invention relates to a method and apparatus for measuring the optical state on the inside of a reaction vessel. The apparatus includes: linking portions that optically connect with the interior of the reaction vessel that is linkable with reaction vessels, a connecting end arranging body having an arranging surface that arranges and supports along a predetermined path, connecting ends to which is provided back ends of the light guide portions, the front ends thereof being provided to the linking portions, the connecting ends being provided to the linking portions; a measurement device having measuring ends that are successively optically connectable with the connecting ends, and in which light based on the optical state is receivable by optical connections between the connecting ends and the measuring ends; and a light guide switching mechanism that successively optically connects the connecting ends and the measuring ends.

CROSS REFERENCE

This application is a continuation of U.S. patent application Ser. No.14/415,072, which is a United States national phase application ofco-pending international patent application number PCT/JP2013/069385,filed Jul. 17, 2013, which claims priority to Japanese patentapplication number 2012-159088, filed Jul. 17, 2012, the entiredisclosures of which are hereby incorporated herein by reference.

TECHNICAL FIELD

The present invention relates to a light measurement apparatus for areaction vessel, and a light measurement method therefor.

BACKGROUND ART

At the time reactions such as amplification of nucleic acids (DNA, RNA,and the like) and the fragments thereof (oligonucleotides, nucleotides,and the like) are performed, in tests that require quantitativeness,such as the analysis of gene expression levels, it becomes necessary toperform the amplification such that the ratio of the relative amounts ofthe respective nucleic acids can be known. Consequently, by using thereal-time PCR method, and by using an apparatus provided with a thermalcycler and a fluorescence spectrophotometer, analysis by electrophoresisis made unnecessary as a result of the generation process of the DNAamplification products in PCR being detected and analyzed in real time.Furthermore, as a DNA amplification method that performs amplificationwhile maintaining the quantitativeness with respect to the ratio of therelative amounts of the DNA or RNA contained in the sample beforeamplification, the SPIA (Single Primer Isothermal Amplification) methodis used. In the SPIA method, the linear DNA amplification methodresulting from an isothermal reaction utilizing DNA/RNA chimera primer,DNA polymerase, and RNaseH has become used.

In a case where processing such as nucleic acid amplification, andmeasurements thereof are performed, conventionally, the target nucleicacid is separated and extracted from the sample by using a filter bymeans of a manual method, by using magnetic particles and adsorption onan inner wall of a vessel or a pipette tip by means of a magnetic field,or by using a centrifuge. The separated and extracted target compound istransferred and introduced into a reaction vessel together with areaction solution by a manual method and the like, and upon sealing ofthe reaction vessel using a manual method and the like, at the timereactions are performed using a temperature control device forreactions, optical measurements are performed with respect to thereaction vessel using a light measuring device (Patent Document 1).

In a case where the processing is executed by a manual method, a largeburden is forced on the user. Furthermore, in a case where theprocessing is executed by combining a dispenser, a centrifuge, amagnetic force device, a temperature controller, a device for sealingthe reaction vessel, a light measurement device, and the like, there isa concern of the scale of the utilized devices increasing and of thework area expanding. In particular, in a case where a plurality ofsamples is handled, since it becomes necessary to separate and extract aplurality of target nucleic acids and for amplification to be torespectively performed, the labor thereof becomes even greater, andfurthermore, there is a concern of the work area also expanding further.

Specifically, in a case where reactions of the nucleic acids (DNA, RNA,and the like) to be amplified, and the like, are performed within aplurality of reaction vessels and these reactions are monitored byoptical measurements, the measurements are performed by successivelymoving a single measuring device to the respective reaction vessels by amanual method, or the measurements are performed by providing ameasuring device to each of the respective reaction vessels beforehand.

In the former case where a single measuring device is used, when themeasuring device is attempted to be manually moved to the apertures ofthe reaction vessels, there is a concern of subtle differences occurringin the measurement conditions for each reaction vessel as a result ofsubtle displacements or relative motions between the reaction vessel andthe measuring device.

In the latter case where a measuring device is provided to each of therespective reaction vessels, although the positioning accuracy becomeshigh, there is a concern of the devices scale expanding, and of themanufacturing costs increasing. Furthermore, although it is preferableto seal the apertures of the reaction vessels at the time of temperaturecontrol and the measurements, it is time-consuming to perform sealing,or opening and closing, with respect to a plurality of reaction vesselsby a manual method with a lid, and in particular, there is a concern ofthe lid becoming adhered to the vessel apertures such that it becomesdifficult to easily open the lid, and of contamination occurring fromthe liquid attached to the inside of the lid dripping or splashing.Furthermore, there is a concern of providing a dedicated opening andclosing mechanism of the lid, complicating the apparatus, and increasingthe manufacturing costs (Patent Document 2).

As a device that automatically performs measurements without providing ameasurement device for each of the respective reaction vessels, there isan device that, at the time temperature control of a microplate having aplurality of wells is performed by a thermal cycler, successive lightmeasurements of the respective wells is performed by moving a detectionmodule over the microplate (Patent Documents 3 and 4).

In this device, since the detection module itself is moved in a state inwhich it is supported by the thermal cycler, a load that accompanies theacceleration from the movement is imparted on the detection module,which has precision optical system elements and electronic circuits suchas a photomultiplier tube, thus becoming the cause of noise orbreakdowns of the measurement device, and furthermore, there is aconcern of the device lifetime being shortened.

Moreover, since the detection module is supported by the microplate, oris supported by a lid body sealing the respective wells of themicroplate and only moves in a horizontal direction, a fixed spacingbetween the respective wells and the measuring end of the measurementdevice is necessary. Therefore, since attenuation from the scattering oflight, and the leakage or entry of light with respect to the adjacentwells cannot be completely blocked and prevented, there is a concern ofa measurement with a high accuracy not being able to be performed.

Furthermore, since the detection module divides the light path using ahalf mirror at the time light from the vessels is received or isirradiated, there is a need to take a long light path length within themeasurement device, and it has a problem in that there is a concern ofthe device scale becoming large.

In a case where the detection module is one that moves such that itpasses the respective wells that are arranged on the microplate and thenumber of wells becomes large, there is a concern of the processing timebecoming long due to the movement distance being long, and in addition,there is also a concern of the problems of the measurement devicementioned above occurring.

Moreover, at the time an optical measurement is performed on a sealedreaction vessel, there is a concern of the lid which has transparency,or the optical system elements, becoming cloudy from condensation, andthe measurements becoming difficult.

Consequently, in order to perform nucleic acid amplification and thelike, as a precondition thereof, specialized researchers or techniciansbecome necessary, and this situation is preventing the generalization ofgenetic analysis and the expansion of clinical applications inhospitals, and the like.

Therefore, at the time of clinical use and the like, in order to preventcross-contamination and to reduce user labor, and to easily perform thegenetic analysis of nucleic acids from the extraction, theamplification, and further, by means of a measurement, then consistentlyautomating the steps from extraction of the target compound, reactingsuch as amplifying, up to measurement, and miniaturization of thedevice, and the provision of an inexpensive, high-accuracy device areimportant.

PRIOR ART DOCUMENTS Patent Documents

[Patent Document 1] International Publication WO96/29602

[Patent Document 2] Japanese Unexamined Patent Publication No.2002-10777

[Patent Document 3] United States Patent No. 7148043

[Patent Document 4] United States Patent No. 7749736

SUMMARY OF THE INVENTION Problems to be Solved by the Invention

Therefore, the present invention is one that has been achieved in orderto solve the problems mentioned above. A first object thereof is inproviding a light measurement apparatus for a reaction vessel, and amethod thereof that, with regard to the optical state within a reactionvessel with respect to nucleic acids and the like, makes rapid andefficient measurements with a high accuracy and reliability possible.

A second object is in providing a light apparatus for a reaction vessel,and a method thereof that, by simplifying the construction of theoptical system and performing measurements using a small number ofmeasurement devices with respect to a plurality of reaction vessels,prevents the expansion of the apparatus scale and increases in thecomplexity of the apparatus construction, and can be inexpensivelymanufactured and utilized.

A third object is in providing a light measurement apparatus for areaction vessel, and a method thereof that, by consistently automatingin parallel the optical measurements with respect to the plurality ofreaction vessels, in which reactions such as amplification of nucleicacids are performed, and the associated processing therein, processingwith a high reliability can be performed by preventing with certainty;contaminations due to the entry of contaminants into the plurality ofreaction vessels from the exterior, or liquid leakage from the pluralityof reaction vessels, and the cross talk of light between adjacentmeasurement devices at the time of measurement.

Means for Solving the Problem

A first aspect of the invention is a light measurement apparatus for areaction vessel comprising: two or more linking portions to which isprovided front ends of one or two or more light guide portions that aredirectly or indirectly linkable with two or more reaction vessels andoptically connect with the interior of the reaction vessel to which itis linked;

a connecting end arranging body that has an arranging surface thatarranges and supports along a predetermined path, two or more connectingends, to which is provided back ends of the light guide portions, inwhich the front ends thereof are provided to the linking portions, theconnecting ends are provided corresponding to the respective linkingportions;

a measurement device that has one or two or more measuring ends that aresuccessively optically connectable with the respective connecting endsalong a predetermined path, and in which light based on an optical statewithin the reaction vessels is receivable by means of opticalconnections between the connecting ends and the measuring ends, themeasuring ends are provided approaching or making contact with thearranging surface; and a light guide switching mechanism that relativelymoves the respective connecting ends arranged on the connecting endarranging body and the respective measuring ends such that they aresuccessively optically connected. In a case where the measurement devicecomprises a plurality of types of measurement devices, it is preferableto provide with respect to the respective measurement devices, and atthe very least between adjacent measurement devices, a modulator thatmodulates the intensity of the light to be received by the measurementdevices at mutually different predetermined frequencies, and a tunerthat, with regard to the light received by the measurement devices,performs tuning to the predetermined frequencies, and obtains thecorresponding intensity of the received light.

It is preferable for the vessel group, to which the reaction vessels areprovided, to have in addition to the reaction vessels, two or moreliquid housing parts that house liquids such as samples, reagents, andthe like. Furthermore, the vessel group includes a microplate in whichwells representing a plurality of liquid housing parts are arranged in amatrix form or a column (row) form, or a cartridge form vessel in whichwells representing a plurality of liquid housing parts are arranged in arow form. In a case where amplification of nucleic acids is performed,the vessel group is provided with two or more liquid housing partshousing for example; a sample, a magnetic particle suspension in whichmagnetic particles that are able to capture the nucleic acids or thefragments thereof, which represent the amplification subject, aresuspended, a solution for separating and extracting used for theseparation and the extraction of the amplification subject, and anamplification solution used in nucleic acid amplification.

Furthermore, it is preferable for the two or more linking portions to beprovided such that they are arranged on the light guide stage.

Here, the “amplification solution” represents, in a case whereamplification is performed by the PCR method for example, a template DNAsolution which is the amplification subject, a primer solution, a DNApolymerase solution, a nucleotide solution, a reaction buffer solution,and the like. In a case where amplification is performed by the SPIAmethod, it represents a DNA/RNA chimera primer solution, a DNApolymerase solution, an RNaseH solution, and the like. Furthermore,generally, methods for performing real-time PCR using fluorescentreagents containing a fluorescent compound include the intercalationmethod, the hybridization method, and the LUX method. In the“intercalation method”, a fluorescent compound such as SYBR (registeredtrademark) GREEN I or ethidium bromide, enters into double-stranded DNAat the time of the elongation reaction, and is a method in which the DNAamount is measured by irradiating an excitation light and utilizing thefluorescent light-emitting characteristics. Therefore, at the veryleast, the fluorescent material and a quencher that suppresses the lightemission of the fluorescent material must be contained within theamplification solution. The “hybridization method” is a method thatdetects only a target PCR product by using a DNA probe labeled with afluorescent material in addition to a PCR primer. That is to say, as aresult of the DNA probe labeled by the fluorescent light fluorescentmaterial hybridizing with the target PCR product, the hybridized DNA(amount) thereof is detected. The “LUX method” is one that utilizes aproperty in which the fluorescent light signal of the fluorescentcompound labeling the oligonucleotide is affected by the shape (such asa sequence, a single-strand, or a double-strand) of the oligonucleotidethereof. In actual real-time PCR, a PCR primer (LUX primer) that islabeled with one type of a fluorescent compound and a contrastinglyunlabeled PCR primer are used to perform real-time PCR. The LUX primerthereof is labeled with a fluorescent compound in the vicinity of the3′-terminus, and is designed such that it takes a hairpin structure inthe interval between the 5′-terminus. When the LUX primer takes ahairpin structure, the quenching effect is resolved, and the fluorescentlight signal becomes increased. By measuring this signal increase, theamount of the PCR product can be measured.

Examples of the material of the vessels, which includes the reactionvessels, the lid, and the like, include resins such as polyethylene,polypropylene, polystyrene and acrylic, glass, metals, and metalcompounds. The size of the vessels is, in addition to several μL toseveral 100 μL of liquid being storable, a size in which the ends of thedispensing tips are insertable for example. In the case of a cylindricalshape, the diameter of the size of one vessel is several mm to several10 mm, and the depth is several mm to several 10 mm for example.

It is preferable for the interior of the reaction vessels to betemperature controllable by a temperature controller.

The “temperature controller” has a temperature source that is able tolower the temperature within the reaction vessels, which house theliquids that become subjected to temperature control, based on a signalfrom the exterior for example. The temperature source is one in which,for example, a Peltier element, a heater, a cooling device, and the likeis provided on a block-shaped member. In order to perform processingsuch as PCR, the temperature controller is preferably a thermal cyclerusing a Peltier element. That is to say, it is preferable fortemperature control to be achieved by providing a block for temperaturecontrol as a temperature source to the vessel group or on the stage, inwhich temperature is raised and lowered by means of a Peltier element,that makes contact with or is adjacent to a portion (a lower side wallsection for example) or the entirety of the reaction vessel.Furthermore, it is also possible to perform temperature control of anisothermal amplification by means of the LAMP method.

“Temperature control” represents, with respect to a liquid or a vesselthat becomes the subject thereof, the maintaining of one or two or moreset predetermined temperatures for set time periods, according to aspecified sequence, and the execution at a specified frequency. Theinstructions to the temperature controller are carried out by sending acorresponding signal based on a program.

The “predetermined temperature” is a target temperature that an object,such as a liquid that becomes the subject, is to reach. In a case wherenucleic acids, such as the DNA contained in a liquid, oroligonucleotides and the like, which represent fragments of nucleicacids, are amplified by the PCR method for example, the predeterminedtemperature that is set is a temperature cycle performed in the PCRmethod. That is to say, it represents temperatures that are respectivelynecessary for the denaturation, the annealing or the hybridization, andthe elongation of DNA of approximately 94° C., a temperature in theinterval from 50° C. to 60° C., and a temperature of approximately 72°C. for example. On the other hand, in the case of the SPIA method(trademark), it becomes set at a fixed temperature, such as 55° C. forexample.

Furthermore, the predetermined temperature includes a temperature fortransition acceleration that shortens the transition time and keeps thesingle cycle time within a predetermined cycle time as a result of, inthe case of a transition from a high-temperature predeterminedtemperature to a low-temperature predetermined temperature, performingcooling at a temperature for transition acceleration that is lower thanthese predetermined temperatures by means of the temperature controller,or, at the time of a transition from a low-temperature predeterminedtemperature to a high-temperature predetermined temperature, byperforming heating at a temperature for transition acceleration that iseven higher than these predetermined temperatures for example. The“predetermined time” is the time necessary for maintaining therespective temperatures, and although it depends on the type of theamplification method, the reagents and the amount of liquid used in thePCR method, and the shape, the material, the size, the thickness, andthe like, of the nozzles, a single cycle is, in total, from severalseconds to several 10 seconds for example, and the processing time forthe PCR method as a whole is of the order of approximately severalminutes to several 10 minutes for example. The transition time is alsoincluded in the predetermined time.

The “linking portion” is a member that is able to be releasably linkedwith the reaction vessel directly, or indirectly via the sealing lid andthe like. Provided to the linking portion is the end of a light guideportion that is able to guide the light based on the optical statewithin the reaction vessel, by optically connecting with the interior ofthe reaction vessel. Here, the “linking with the reaction vessel”represents approaching or joining with the aperture, the outer wall, orthe outer bottom portion of the reaction vessel or a mounted sealing lidor sleeve and the like. Furthermore, “approaching” represents, withoutmaking contact, an approach to an extent that optical connection of theinterval with the light guide portions is possible. Moreover, “joining”includes making contact, close contact, adhesion, fitting, and mounting,and at the very least represents making contact such that opticalconnection of the interval between the light guide portions is possible.As a result of this linking, the light guide portion provided to thelinking portion and the interior of the reaction vessel are opticallyconnected. The linking portion is a plate-shaped section of the lightguide stage mentioned below to which the light guide portion is providedfor example. The “light guide portion” is an optical system element, ora combination thereof, in which light is able to pass through. The endof the light guide portion is a hole piercingly provided in theplate-shaped section thereof, a transparent section such as an opticalfiber, or an optical system element such as a lens. Alternatively, forexample, it is a member of a cylindrical shape, and the like, providedsuch that it protrudes from the light guide stage, and the end of thelight guide portion is a cavity provided in the member of a cylindricalshape, and the like, a transparent section such as an optical fiber, oran optical system element such as a lens, or one end thereof. It ispreferable for the light guide portion to be such that a portion thereofhas a flexibility. In this case, it represents an optical fiber or anoptical fiber bundle, or a combination of these with an optical elementsuch as a lens for example. If fluorescent light is measured, there is acase where two or more light guide portions are provided, and in such acase, they can be used such that a portion thereof are for irradiation,and the others are for receiving light. The connecting ends are providedwith these optical elements, or the other ends thereof. Furthermore, acase where it is directly linked with the aperture of the reactionvessel represents a case in which the interior of the reaction vessel issealed using mineral oil and the like, and in this case, it ispreferable to form the linking portion such that it is able to directlyseal the reaction vessel. Furthermore, in a case where the linking isperformed outside of the aperture, there is a need for the reactionvessel or the linking section thereof to have a transparency. It ispreferable, in order to prevent condensation, for the material of thelinking portion to be formed by a resin such as a PEEK material, whichhas a low heat conductivity and has a rigidity, such that the heat fromthe heating portion is not released through the linking portions.Furthermore, if the heat from the heating portion is not releasedthrough the linking portions, it is possible to prevent the effects ofheat toward the optical system elements, such as an optical fiber or alens, that are provided to the linking portion. The spacing betweenadjacent connecting ends on the arranging surface of the connecting endarranging body is preferably formed such that it becomes smaller thanthe spacing between adjacent linking portions on the light guide stage.Consequently, it becomes possible integrate the arrangement of theconnecting ends with respect to the arrangement of the linking portions.

The “predetermined path” represents, as a result of the measuring endsand the connecting end arranging body relatively moving, a path on aplane surface or a curved surface whereby the measuring ends are able toscan all of the connecting ends arranged therealong. Furthermore, thepath that connects all of the connecting ends represents a single ormultiple line segments that do not intersect (including zigzag lines andclosed straight lines), curved lines (including spirals and closedcurved lines), or a path along a combination of these and the like.Preferably, the respective single or multiple paths are continuous, andpaths along straight line segments without cusps or corners, or smoothcurves that have a curvature that the measuring ends are able to follow,are preferable.

There is a case where the linking portions and the connecting endscorrespond one-to-one, a case where they correspond many-to-one, and acase where they correspond one-to-many. Here, midway, it is possible forthe light guide portions to be branched or joined, or a light guideportion bundle comprising a plurality of light guide portions to bebranched or joined.

It is preferable for the predetermined path to be determined based onthe number, the shape, the arrangement, or the size of the measuringends on the measuring device, such that smooth scanning is possible. Forexample, a predetermined path along a straight line in which, for themovement of the connecting ends with respect to the measuring ends,there are no sudden changes in direction, such as changes to an obtuseangle or a right angle direction with respect to the travelingdirection, is preferable.

The arrangement pattern of the linking portions is a matrix form, acolumn form, or a row form for example. The arrangement pattern of theconnecting ends is the same arrangement, or a similar arrangement thatdiffers only in size for example, or in a case where the arrangementpattern is different, examples include the case of a circular form,other closed curved forms, a single column form, or a matrix form havinga smaller number of columns or rows. The predetermined path isdetermined such that it passes through all of the arranged connectingends. The connecting ends also include a case where they are mutuallymovably provided with respect to the linking portions, and a case wherethey are immobile with respect to the linking portions, and are providedas a connecting end arranging body on a connecting end arranging surfaceon the opposite side of the arranging surface of the linking portions ofthe same light guide stage.

Furthermore, it is preferable for the arrangement of the connecting endsto be such that they are integrated with respect to the arrangement ofthe linking portions. In this case, it is preferable for the light guideportion to have a flexibility. “Integration” is preferably performed bymeans of the predetermined path (or the arrangement pattern of theconnecting ends) representing a smaller region area or a smaller spacingthan the region area that encloses the arrangement pattern of thelinking portions on the light guide stage or the spacing betweenadjacent linking portions, and by making the total scanning distanceshort. Consequently, in a case where the speed is made the same,processing within a shorter time than a case where the linking portionsdirectly scan the measuring ends is possible. Furthermore, the scale ofthe apparatus is also made spatially smaller, which is of benefit forreducing manufacturing costs and the effective use of a workspace.

For example, in a case where “integration” is performed by making thespacing of adjacent connecting ends on the arranging surface of theconnecting end arranging body smaller than the spacing of adjacentlinking portions on the light guide stage, it was experimentallyconfirmed that when the plurality of reaction vessels were arranged witha spacing of 9 mm pitch, and when the external diameter of the opticalfiber of the light guide portion was 1.5 mm, the pitch of thearrangement of the connecting ends could be set to 3.75 mm.

The extent of integration is such that the greater it becomes, the morepreferable it is. This is because the processing time is shortened andthe scale of the apparatus is spatially compressed. However, at the veryleast, it is at most an extent in which the relative movement orscanning of the connecting end arranging body and the measuring deviceis able to complete the receiving of the light from all of the reactionvessels to be measured within the stable light receivable time. Here,the “stable light receivable time” represents the time in which theoptical state within the reaction vessels, for which the light isreceivable, is stably maintained. In the case of the intercalationmethod or the LUX method of real-time PCR, or the TaqMan probe of thehybridization method for example, it corresponds to the time in whichthe elongation reaction of the respective cycles of PCR is performed. Ina case where a FRET probe is used in the hybridization method, itcorresponds to the time in which annealing is performed.

Consequently, it can be applied with respect to a light emitter with ashort stable light receivable time and the like, and the versatility ishigh.

If the time taken for a single cycle is made several 10 seconds orseveral minutes for example, the stable light receivable time becomesseveral seconds or 10 seconds. However, the fluorescent light detectionamount of the initial cycles of a PCR reaction is below the detectionlimit, and the later cycles of the PCR reaction become a plateau state,and in order to secure quantitativeness by a strict definition, it mustbe within a range of the amplification curve in which an exponential PCRamplification can be observed. The present invention is one in which thestable light receivable time utilizes the fact that the movement time ofthe measuring end between the reaction vessels can be used, and byperforming the relative movement necessary for receiving the light fromthe respective reaction vessels within the stable light receivable time,the receiving of the light from the plurality of reaction vessels can beperformed approximately in parallel by means of a single measuringdevice, or a sufficiently small number in comparison to the number ofreaction vessels, without using complicated optical system elements andwithout expanding the scale of the apparatus.

The “optical state” represents a state such as light emissions, colors,color changes, or light variations. The light based on the optical staterepresents light from light emissions or light variations, or reflectedlight from light irradiated with respect to colors or color changes, ortransmitted light, scattered light and the like.

The “connecting ends and the measuring ends are successively opticallyconnected” represents that the connecting ends and the measuring endsare optically connected by becoming opposed at a close proximity. Sincethe amount of light received by the measuring device at the moment ofconnection corresponds to a maximum value, the measurement controlportion specifies the data to be measured by calculating the maximumvalue of the amount of light.

The “measuring device” is one that makes fluorescence andchemiluminescence measurements possible for example, and in the formercase, it has a filter for the irradiation of one or two or more types ofexcitation light and the receiving of fluorescent light having one ortwo or more types of wavelengths. It is preferable for these to beguided using an optical fiber. Furthermore, there is a case where theyare provided to the plurality of types of measurement devices such thatthey correspond to the magnitude and the range of wavelengths (orfrequencies).

The “measuring end” has, at the very least, an inlet for the light to bereceived provided to the measuring device, and in the case of afluorescence measurement, has an outlet for the light to be irradiated.These can be provided as separate measuring ends. The inlet and theoutlet are respectively optically connected to a light receiving portioncomprising a photoelectric element provided in the interior, or to anirradiation source. At that time, they can be respectively connected viathe light guide portion for receiving light and the light guide portionfor irradiation. Furthermore, the connecting end arranging body, themeasuring ends, and the measurement device are preferably provided atseparated locations such that they do not directly make contact or arenot in the vicinity of the reaction vessels or the stage for mounting,in which heating control or temperature control is performed.

The light measurement apparatus for a reaction vessel, although notexplicitly specified, additionally has “a measurement control portion”.The “measurement control portion” controls the measuring device and alight guide switching mechanism, comprises a computer (CPU) built intothe light measurement apparatus for a reaction vessel and a program thatdrives the computer, and achieves measurement control by transmitting asignal through a DA converter to the respective control portions thatdrive the transfer mechanisms for example.

Here, the “modulator” represents a device that modulates the strength ofthe light to be received by the respective measurement devices atpredetermined frequencies. For example, in a case where the opticalstate within the reaction vessels represents the emission of fluorescentlight, it has an excitation light modulation portion as described below.The excitation light modulation portion includes a drive circuit, suchas an oscillating voltage supply circuit that makes a voltage applied toan irradiation source, which is for generating an excitation light to beirradiated to a fluorescent compound, an oscillating voltage having apredetermined frequency, and various devices for blinking of the lightto be received at the predetermined frequency, such as an opticalshutter, a rotating polygon mirror, and an optical chopper formed by alight transmitting-hole type rotating body. Examples of the “oscillatingvoltage of the predetermined frequency” include sinusoidal oscillation,and pulse wave-type variations including blinking of the light. The“tuner” represents a device that, with respect to the received light,obtains the corresponding light intensity of the received light byperforming tuning to the predetermined frequency. For example, it has abandpass filter circuit that extracts the frequency or a frequency band.This is generally because the wavelength of the excitation light isshorter than the wavelength of the fluorescent light, and the intensityis higher. Therefore, it is possible that measurements are performedwhere a plurality of types of fluorescent light and a plurality of typesof excitation light are used in a state where they are mutually similar.In such a case, if the wavelength of a given excitation light and thewavelength of the fluorescent light overlap, the extraction of thefluorescent light from the excitation light becomes difficult.Therefore, at the very least between adjacent measurement devices, theycan both be discriminated by performing modulation to differentpredetermined frequencies. Consequently, other optical noise, such asnatural light, can be excluded.

The “predetermined frequency” is in a range from 1 Hz to 1 MHz, and ispreferably from approximately 1 kHz to 10 kHz. Since it is “at the veryleast between adjacent measurement devices”, it is possible to use thesame frequency between measurement devices that are separated by a fixeddistance or more.

A second aspect of the invention is a light measurement apparatus for areaction vessel in which, at the time light is received by themeasurement device, at the very least, a measurement device bodyexcluding the measuring ends is immovably provided with respect to thelight guide stage, which is provided with the reaction vessels and thelinking portions linked with the same.

Therefore, there is a case where the connecting end arranging body moveswith respect to the measuring ends, or the measuring ends move withrespect to the connecting end arranging body, and the measurement devicebody may be movably provided with respect to the reaction vessels or thelight guide stage until the light guide stage is linked with thereaction vessels. The former case represents a case where themeasurement device body is linked with the light guide stage or a casewhere it is linked with movements in a portion of the directions forexample. Furthermore, the latter case represents a case where themeasurement device body is linked with the reaction vessels, or is fixedto the stage together with the reaction vessels. The measuring ends, ina case where they are present, include light guide portions, which areon the exterior of the measurement device body, up to the measuringends.

A third aspect of the invention is a light measurement apparatus for areaction vessel, wherein the measurement device comprises a plurality oftypes of specific wavelength measurement devices that are able toreceive light of specific wavelengths or specific wavelength bands, thespecific wavelength measurement devices having at least one measuringend that is successively optically connectable with the connecting endsalong the predetermined path, and the frequencies of the modulatorsprovided to the specific wavelength measurement devices being mutuallydifferent, at least between adjacent predetermined specific wavelengthmeasurement devices. In a case where the light based on the opticalstate is a fluorescent light, the specific wavelength measurementdevices are provided with an irradiation source that irradiates apredetermined excitation light that excites the correspondingfluorescent light of a specific wavelength or a specific wavelengthband, and a light receiving portion that is able to receive thefluorescent light of the specific wavelength or the specific wavelengthband, and the modulators that correspond to the respective specificwavelength measurement devices are provided with an excitation lightmodulation portion that modulates the excitation light at thepredetermined frequency, and the tuners are provided with a bandpassfilter circuit or a lock-in amplifier that, with respect to the lightreceived by the light receiving portion, extracts the intensity data ofthe light having the predetermined frequencies.

Here, the measuring end is provided with a cavity, an optical elementsuch as a lens, or a light guide portion such as an optical fiber forexample. Furthermore, the measuring end is provided with an irradiationaperture that connects with the irradiation source, and a lightreceiving aperture that connects with the light receiving portion. The“bandpass filter circuit” represents a circuit that extracts from ameasurement subject signal only the light signal that possesses afrequency band that contains the (modulated) predetermined frequency,and removes signals of all other frequencies. Furthermore, a filtercircuit in which a high-pass filter and a low-pass filter are combinedis exemplified. The “lock-in amplifier” represents a circuit that takesa measurement subject signal and a reference signal having the modulatedpredetermined frequency, multiplies the two by means of a multiplier(Phase Sensitive Detector), and by smoothing thereafter, obtains anoutput signal possessing the intensity component of the signal havingthe predetermined frequency.

A fourth aspect of the invention is a light measurement apparatus for areaction vessel, that has two or more linking portions provided on thelight guide stage, and a stage transfer mechanism that relatively movesthe light guide stage with respect to the reaction vessels such that thelinking portion is simultaneously directly or indirectly linked with thetwo or more reaction vessels.

Preferably, the spacing between adjacent connecting ends on thearranging surface of the connecting end arranging body is formed suchthat it becomes smaller than the spacing between adjacent linkingportions on light guide stage.

In a case where the stage transfer mechanism makes the light guide stagerelatively movable in the vertical direction with respect to the vesselgroup, the sealing lids that are mounted such that they cover theapertures of the reaction vessels can be pressed or shaken. That is tosay, it is preferable for the measurement control portion, followingindirectly linking with the linking portions via the sealing lids suchthat it covers the apertures of the reaction vessels, to perform controlsuch that it presses or shakes the sealing lids. By pressing, thesealing of the reaction vessels can be performed with certainty. Inaddition, by shaking, the sealed state between the apertures of thereaction vessels and the sealing lids can be rapidly and easily removedand released. Therefore, a high processing efficiency and reliabilitycan be obtained.

In a case where the linking portions are not linked by joining, such asby directly or indirectly fitting with the apertures of the reactionvessels, but by linking with the reaction vessels by approaching thereaction vessels, it is possible to successively and smoothly repeat thelinking between the linking portions and the reaction vessels, and thereleasing thereof, by means of horizontal movements without performingrelative movements in the vertical direction.

Furthermore, the two or more linking portions provided to the lightguide stage are arranged on a linking portion arranging body that ismovable in the horizontal direction with respect to the light guidestage in a state in which they are directly or indirectly simultaneouslylinkable with two or more reaction vessels. Moreover, by moving thelinking portion arranging body with respect to the light guide stage, alight measurement apparatus for a reaction vessel that, without movingthe light guide stage, makes more reaction vessels linkable than thenumber of reaction vessels that are simultaneously linkable by thelinking portion arranging body, can be provided. In this case, it ispreferable for the linking between the respective linking portions andthe reaction vessels to be performed within shielded regions that aremutually shielded, such as within two or more grooves or two or moreregions that are mutually separated by a partition wall, to which therespective linking portions are insertable, that extend in a horizontaldirection in which the linking portion arranging body thereof ismovable, and are also provided to the light guide stage for each of thelinking portions. Consequently, the contamination of light from otherreaction vessels can be prevented with certainty.

In this case, the linking portions can be easily and rapidly linked withthe reaction vessels by merely moving in the horizontal directionwithout movements of the light guide stage in the vertical direction.Therefore, by setting the speed of the linking portion arranging bodysuch that it includes the horizontal movements of the linking portionarranging body and can be performed within the stable light receivabletime, it becomes possible to receive light and perform measurementsessentially in parallel with respect to an even greater number ofreaction vessels by means of a single set of measurement devices.

In a case where the measurement device has one or two or more measuringends that are optically connectable with the connecting ends, and aplurality of types specific wavelength measurement devices that are ableto receive light of specific wavelengths or specific wavelength bands,then preferably provided is a measuring end aligning portion that alignsthe plurality of measuring ends such that they are optically connectablewith the respective connecting ends along the predetermined path.

The “aligning” is integrally or linkingly performed. “Integrally”represents arrangement such that the intervals between the measuringends are mutually fixed and do not have any degrees of freedom.“Linkingly” represents arrangement such that the intervals between themeasuring ends have degrees of freedom to some extent, such as in achain. There is a case where the “aligning” is such that the respectivemeasuring ends are lined up along the scanning direction of thepredetermined path, or a direction perpendicular to the scanningdirection. In the latter case, the predetermined path is such that aplurality of paths are lined up in parallel.

According to the present aspect of the invention, by using a pluralityof types of luminescent compounds, colored compounds, color changingcompounds, or light variation compounds, and performing amplificationprocessing in parallel under the same conditions on a plurality of typesof amplification subjects in a single reaction vessel, it is possible toperform multiplex PCR amplification or multiplex real-time PCR on aplurality of types of amplification subjects by using a primer labeledwith a plurality of types of luminescent compounds for example.

Since it is “light of a specific wavelength or a specific wavelengthband”, it represents, in terms of visible light, a range of wavelengthssuch as a red light, a yellow light, a green light, a blue light or aviolet light for example.

A fifth aspect of the invention is a light measurement apparatus for areaction vessel wherein the vessel group is provided with sealing lids,which have transparency, that are mounted on apertures of one or two ormore of the reaction vessels, and seal the reaction vessels.

Here, the “sealing lid” includes, in addition to those that areinflexible and a plate form or block form, those that are a film form ora membrane form and have a flexibility. The “mounting” includes fitting,threading, friction, adsorption, attachment, adhesion, and the like. Inthese cases, detachable mounting is preferable.

Furthermore, in a case where the respective linking portions of thelight guide stage are linked at the apertures of the respective reactionvessels, it is preferable to make the linking portions or the nozzlespressable or shakable with respect to the sealing lids covering theapertures of the reaction vessels.

It is preferable for the linking portions to be provided such that theydownwardly protrude from the light guide stage. In this case, thelinking portions, for example, have a shape such as a rod shape, acylinder shape, a cone shape, and the like, and the lower end portionsof the members are preferably able to make contact with the sealinglids.

The sealing lids singularly cover the apertures of one or two or morereaction vessels. The sealing lids are moved by being mounted to thenozzles mentioned below, and cover the apertures of the reaction vesselsby using the tip detaching mechanism for example. To accomplish this,one or two or more indentations for mounting that are mountable on theone or two or more nozzles are provided on the upper side of the sealinglids. The one or two or more linking portions can be linked with thereaction vessels by being inserted within these indentations (which arealso indentations for linking) by means of vertical direction movementsof the light guide stage.

It also is possible, without moving the sealing lids by means of thenozzles, to provide a dedicated sealing lid transport mechanism. As thesealing lid transport mechanism, the light measurement apparatus for areaction vessel has a transporting body that is movable with respect tothe vessel group, and one or two or more grippers arranged on thetransporting body according to the arrangement of the reaction vesselsthat, with respect to the cover plate that covers the apertures of therespective reaction vessels, and the sealing lids having mountingportions that protrude on the lower side of the cover plate excludingthe center portion, in which light is able to pass through, and to whichthe cover plate is mountable to the reaction vessel, grips the coverplate such that the mounting portions are exposed on the lower side in astate in which they are mountable to the reaction vessels for example.Furthermore, if the sealing lid transporting body is made to be linkedwith the light guide stage, the apparatus construction is simplified,and the expansion of the apparatus scale can be prevented.

In this case, since it is not necessary to provide an indentation formounting the nozzles and the like on the upper side of the sealing lid,the linking portion can be easily linked by just horizontal directionmovement between the apertures of the reaction vessels on the sealinglid without vertical direction movements of the light guide stage. Inthis case, if the horizontal direction movement of the linking portioncan be performed within the stable light receivable time, it becomespossible to receive light and perform measurements essentially inparallel with respect to an even greater number of reaction vessels.

A sixth aspect of the invention is a light measurement apparatus for areaction vessel, wherein the light guide stage is provided with aheating portion that is able to heat the sealing lids.

The measurement control portion, after simultaneously mounting thesealing lids to the linking portions, and following control of the stagetransfer mechanism such that the optical linking portions aresimultaneously indirectly linked with the two or more reaction vessels,controls the heating portion such that the sealing lids are heated forexample. The “heating portion” has a heating function at a temperaturethat is set based on the magnitude of an applied electric current or byan ON/OFF control for example.

Here, the heating of the sealing lids by the heating portion isperformed for preventing condensation at the time of temperature controlof the reaction vessels sealed by the sealing lids.

A seventh aspect of the invention is a light measurement apparatus for areaction vessel provided with a temperature controller having atemperature source that is provided making contact with or approachingthe lower side wall sections of the reaction vessels, and a heatingportion provided such that it is able to make contact with or approachthe upper side wall sections of the reaction vessels positioned furtheron an upper side than the lower side wall sections of the reactionvessels, and that has a heat source that is able to heat the upper sidewall sections.

Here, the “lower side wall section” represents a wall section or aportion thereof including the bottom portion that encloses a volumesection, which is a portion (1% to 90% for example) of the entire volumeof the reaction vessel in which a predetermined liquid amount determinedbeforehand is housable. The lower side wall section represents thesection of the wall section in which the rated liquid amount of liquidis housable for example. In a case where the reaction vessels comprisewide-mouthed piping parts that are linked with the linking portions, anda narrow-mouthed piping part, it is provided on the narrow-mouthedpiping part for example. The “upper side wall section” represents,within the entire volume of the reaction vessel, a vessel sectionenclosing the remaining volume of the lower side vessel section, inwhich the rated liquid amount is housed, or a portion thereof The “upperside wall section” is normally preferably provided on the upper side ofthe reaction vessels leaving a spacing with the lower side wall section.The upper side wall section becomes closer to the aperture, the sealinglid, or the linking portion than the lower side wall section. In thecase of the vessel comprising the wide-mouthed piping part and thenarrow-mouthed piping part, and in a case where the linking portion islinked by fitting with the wide-mouthed piping part, the upper side wallsection is provided on the wall section of the wide-mouthed piping partfor example. The upper side wall section is a section that correspondsto a band shape along the circumference of the vessel wall for example.

The measurement control portion, following controlling the stagetransfer mechanism such that the linking portions are simultaneouslydirectly or indirectly linked with the reaction vessels, controls theheating portion such that direct or indirect condensation on the linkingportions is prevented. “Indirectly linked” represents a case where thelinking portions are linked with the reaction vessels via the sealinglids, the outer walls of the reaction vessels, and the like. “Control ofthe heating portion” is performed according to the “temperature control”for preventing condensation. For example, the heating temperature iscontrolled such that it is set from several degrees (a temperatureexceeding the dew point of water vapor that is necessary for preventingcondensation) to several 10° C. (a temperature that is sufficientlylower than the melting point of the raw material of the reactionvessels), for example, 1° C. to 60° C., or preferably approximately 5°C., higher than the respective predetermined temperatures set by thetemperature control for example. In a case where the amplification is byPCR, heating is performed at a temperature that is several degreeshigher than 94° C., such as 100° C., and in an isothermal case, in acase where the predetermined temperature is approximately 55° C.,heating is performed at a temperature that is several degrees higherthan this for example, such as from approximately 60° C. to 70° C. Theheating temperature depends on the amount of liquid representing thesubject of temperature control, and also the positions of the arrangedreaction vessels. For example, according to experiments, when thetemperature set by temperature control was 95° C. and the amount ofliquid was 25 μL, condensation disappeared at 113° C.

As a result of the heating portion performing heating directly withrespect to the reaction vessels and not the linking portions or thesealing portions, thermal effects toward the optical system elementsprovided to the linking portion, or the measuring ends near the linkingportions, are reduced or removed. Therefore, the degradation of opticalsystem elements such as prisms, optical fibers, various lenses, such asconcavoconvex lenses, ball lenses, aspheric lenses, drum lenses, andgraded index rod lenses, mirrors, waveguide tubes, and the like, isprevented, and the reliability of the image that can be obtained throughthe optical system elements can be increased. For the linking portions,by using various lenses, such as ball lenses and aspheric lenses, whichrepresent the optical system elements, the light that is generatedwithin the reaction vessels and is outgoing in the aperture direction isfocused with certainty, and can be guided by being incident to the lightguide portion, such as an optical fiber.

Here, the reaction vessels; the temperature controller, which has atemperature source provided such that it is making contact with orapproaching the lower side wall sections of the reaction vessels, andperforms temperature control of the interior of the reaction vessels;and the heating portion provided such that it is making contact with orapproaching the upper side wall section, which has a heat source that isable to heat the upper side wall section, configure the reaction vesselcontrol system.

In that case, the reaction vessel preferably comprises a wide-mouthedpiping part, and a narrow-mouthed piping part that is formed narrowerthan the wide-mouthed piping part and is provided on a lower side of thewide-mouthed piping part and communicated with the wide-mouthed pipingpart. The end of the linking portion is fittable to the wide-mouthedpiping part, liquids are housable in the narrow-mouthed piping part, andthe lower side wall section is provided to the narrow-mouthed pipingpart, and the upper side wall section to the wide-mouthed piping part.Furthermore, it is preferable for the contact surfaces between thelinking portion and the upper side wall section of the reaction vessel,which is heated by the heating portion, or the sealing lid which makescontact therewith, to be made as small as possible. Consequently, theeffects of the heating portion toward the optical system elements of thelinking portions can be reduced or removed.

An eighth aspect of the invention is a light measurement apparatus for areaction vessel wherein a linking portion, which is directly orindirectly linkable with the reaction vessels and is provided with anoptical element in the interior, is a material that is usable to atemperature higher than the dew point of water vapor, and has a thermalconductivity smaller than 1.0 W/(m·K) (W: watt, K: kelvin, m: meter).

Here, if the material is a resin for example, in a case where heatingcontrol by the heating portion is performed at 113° C., then as is clearfrom the Tables below, examples of materials that are applicable tothese conditions include PEEK, MC nylon, fluororesins, and PBT. However,when rigidity is considered, PEEK is preferable.

Thermal Properties of Plastics

Coefficient Thermal Continuous usage of linear conductivity temperatureexpansion [W/m · K] [° C.] [1/° C.] PEEK 0.25 −50 to 250 5.0 × 10⁻⁵ MCnylon 0.233 −40 to 120 9.0 × 10⁻⁵ Polyacetal 0.233 −45 to 95  9.0 × 10⁻⁵Fluororesin 0.25 −40 to 250 1.0 × 10⁻⁴ PET 0.24 room temperature to 1005.5 × 10⁻⁵ PBT 0.27 room temperature to 120 1.0 × 10⁻⁴ ABS 0.3 roomtemperature to 50 9.5 × 10⁻⁵ Numerical values are representitive values

A ninth aspect of the invention is a light measurement apparatus for areaction vessel, wherein the light guide stage is provided to a nozzlehead having a suction-discharge mechanism that performs suction anddischarge of gases, and one or two or more nozzles that detachably mountdispensing tips in which the suction and the discharge of liquids ispossible by means of the suction-discharge mechanism, and has a nozzlehead transfer mechanism that makes the nozzle head relatively movablebetween the vessel groups.

In this case, in addition to further providing a magnetic force partthat is able to apply or remove a magnetic force within the dispensingtips mounted on the nozzles or liquid housing parts provided in thevessel group, and which is able to adsorb the magnetic particles on aninner wall of the dispensing tips or the liquid housing parts, it ispreferable to provide an extraction control part that controls thesuction-discharge mechanism, the transfer mechanism, and the magneticforce part, and for the reaction solution, separates and extracts thesolution of the amplification subject from the sample and houses itwithin the liquid housing parts as a portion of the amplificationsolution.

Here, the “solution for separating and extracting” includes a dissolvingsolution that breaks down or dissolves the protein forming the cellwalls and the like contained in the sample and discharges the nucleicacids or the fragments thereof to the outside of the bacteria or thecell, a buffer solution that simplifies the capture of the nucleic acidsor the fragments thereof by the magnetic particles, and additionally, asolution that dissociates from the magnetic particles, the nucleic acidsor the fragments of nucleic acids captured by the magnetic particles. Inorder to perform the separation of the nucleic acids or the fragmentsthereof, it is preferable to repeat the suction and the discharge of themixed solution.

The “dispensing tip” comprises for example a thick diameter portion, anarrow diameter portion, and a transition portion that communicatesbetween the thick diameter portion and the narrow diameter portion. Thethick diameter portion has an aperture for mounting, into which thelower end of the nozzle is inserted and the nozzle is mounted, and thenarrow diameter portion has an end mouth portion in which liquids canflow in and flow out by means of the suction and discharge of gases bythe suction-discharge mechanism. The dispensing tip and the nozzle aremanufactured from organic substances such as resins of polypropylene,polystyrene, polyester, acrylic, and the like, and inorganic substancessuch as glass, ceramics, metals including stainless steel, metalcompounds, and semiconductors.

The “suction-discharge mechanism” is for example a mechanism formed by acylinder, a piston that slides within the cylinder, a nut portion joinedto the piston, a ball screw on which the nut portion is threaded, and amotor that rotatingly drives the ball screw in both forward and reversedirections.

In a case where two or more nozzles are used, by respectively arrangingtwo or more vessel groups so as to correspond to the respective nozzleswithin two or more exclusive regions corresponding to the respectivenozzles, in which a single nozzle enters and the other nozzles do notenter, and by setting the respective exclusive regions for eachdifferent sample, cross-contamination between samples can be preventedwith certainty.

The stage transfer mechanism at least partly utilizes the nozzle headtransfer mechanism. It is preferable for the nozzle transfer mechanism,which moves the nozzles themselves in the Z axis direction, to also atleast partly utilize the nozzle head transfer mechanism, and for thestage transfer mechanism and the nozzle transfer mechanism to beindependently movable with respect to the Z axis direction movement.

A tenth aspect of the invention is a light measurement apparatus for areaction vessel, wherein the nozzle is such that a sealing lid isretainable by mounting, and by detaching the sealing lid, the sealinglid is mountable on an aperture of the reaction vessel. The detaching ofthe sealing lid can be combined with the tip detaching mechanism thatdetaches from the nozzle the dispensing tip mounted on the nozzle. Inthis case, the “sealing lid” has a sealing portion that is mountable onthe aperture of the reaction vessel, and an indentation for linking thatis mountable on the nozzle. Furthermore, in the case of the linkingportion indirectly linking with the reaction vessel by being mounted onthe sealing lid, in a case where the outer diameter of the nozzle andthe outer diameter of the linking portion are different, it ispreferable for the indentation for linking of the sealing lid to befitted with the end of the linking portion in place of the nozzle, bemounted, and make the linking portion able to retain the sealing lid. Inthis case, for the detaching of the sealing lid from the linkingportion, it is preferable to provide a dedicated sealing lid detachingmechanism.

An eleventh aspect of the invention is a light measurement apparatus fora reaction vessel, wherein front ends of a light guide portion bundle,which comprises a plurality of light guide portions, are provided to therespective linking portions, back ends of a light guide portion bundleof a portion of the light guide portion bundle are provided to firstconnecting ends of the connecting end arranging body, a portion or allof the remainder of the light guide portion bundle is provided to secondconnecting ends of the connecting end arranging body, the predeterminedpath comprises a first path and a second path, and by means of movementof the connecting end arranging body, first measuring ends provided onthe measurement device respectively relatively move along a first pathcomprising the first connecting ends, and second measuring ends along asecond path comprising the second connecting ends.

A twelfth aspect of the invention is a light measurement apparatus for areaction vessel, wherein the first measuring end optically connects withan irradiation source of the measurement device, the second measuringend connects with a light receiving portion of the measurement device,and the first measuring end is connectable with the first connectingend, and the second measuring end is connectable with the secondconnecting end.

Here, the connecting end arranging body has a plurality of pairs offirst connecting ends and second connecting ends that are opticallyconnected with the linking portion, and a single pair of a thirdconnecting end and a fourth connecting end that is optically mutuallyconnected by means of the light guide portion. The first measuring endsprovided to the measurement device are relatively movable along a firstpath comprising a plurality of first connecting ends and a single thirdconnecting end, and the second measuring ends provided to themeasurement device are relatively movable along a second path comprisinga plurality of second connecting ends and a single fourth connectingend. The first measuring ends optically connect with the irradiationsource of the measurement device, and the second measuring ends connectwith the light receiving portion of the measurement device. It ispreferable for each pair to be such that the first measuring end issuccessively connectable with the plurality of first connecting ends andthe single third connecting end, and the second measuring ends aresuccessively connectable with the plurality of second connecting endsand the single fourth connecting end.

In this case, the excitation light emitted by the irradiation source isdirectly received by the light receiving portion via the light guideportion without being diverted to the linking portion and the reactionvessel, and the intensity thereof can be measured. Generally, insemiconducting light emitting elements that are used as excitation lightsources, such as LEDs, the emission intensity can change depending onthe temperature. On the other hand, the fluorescence intensity emittedby a fluorescent material dissolved in a solution is proportional to thefluorescent material concentration and the excitation light intensity.Many fluorescence measurements utilize the fluorescent materialconcentration dependence on the fluorescence intensity, and it is ameasurement for estimating the fluorescent material concentration in asolution. However, fluctuations in the excitation light intensity alsochanges the fluorescence intensity in the same manner as fluctuations inthe fluorescent material concentration. Consequently, by measuring theintensity of the excitation light and removing the effects toward thefluorescence intensity based on the fluctuations thereof, ahigh-accuracy measurement can be performed.

Furthermore, it is preferable for the front ends corresponding to thefirst connecting ends and the front ends corresponding to the secondconnecting ends to be arranged such that they are mixed. The “mixing offront ends” is, for a light guide portion bundle comprising a pluralityof light guide portions, preferably an arrangement in which the frontends of two or more types of light guide portions are mixed such thatthey become uniform, such that uniform irradiation, and uniformreceiving of light, are performed.

A thirteenth aspect of the invention is a light measurement apparatusfor a reaction vessel, wherein the vessel group comprises two or moreexclusive regions corresponding to nozzles of respective groups, whichcomprise one or two or more nozzles, in which nozzles of a single groupenter and nozzles of other groups do not enter, and the respectiveexclusive regions at the very least have at least one of the reactionvessels, one or two or more liquid housing parts that house reactionsolutions used in the reactions, and sealing lids that are transportableto the reaction vessels using the nozzles and are able to seal thereaction solutions housed in the reaction vessels, and the light guidestage is extendingly provided across all of the exclusive regions suchthat linking portions of the light guide stage are associated such thatlinking portions of a single group, which comprises one or two or morelinking portions, enter the respective exclusive regions and linkingportions of other groups do not enter.

In order to make “the nozzles of a single group enter and the nozzles ofthe other groups not enter” or “the linking portions of a single groupenter and the linking portions of the other groups not enter”, forexample, this is performed by providing an exclusive region control partthat controls the nozzle head transfer mechanism such that nozzles of asingle group enter the respective exclusive regions and the nozzles ofthe other groups do not enter, and performs control such that thelinking portions of a single group enter the respective exclusiveregions and the linking portions of the other groups do not enter.

A fourteenth aspect of the invention is a light measurement apparatusfor a reaction vessel that is further provided with a set of traversablenozzles comprising one or two or more nozzles that are movable such thatthey traverse the respective exclusive regions, and are able to enterthe respective exclusive regions.

A fifteenth aspect of the invention is a light measurement apparatus fora reaction vessel, wherein inspection information that identifiessamples or shows managed sample information and testing content isvisibly displayed at the respective exclusive regions, and a digitalcamera, which obtains image data by imaging the content displayed at therespective exclusive regions, which includes the sample information andthe testing content, is provided to the traversable nozzles.

Here, the “sample information” represents the information necessary foridentifying or managing the sample, and examples of the information foridentifying the sample include the attributes of the sample, such as thepatient, the animal, the food, the soil, the polluted water, or thelike, from which the sample was collected, and includes the name, theage, the sex, and the ID number of the patient, the sales location ofthe food, and the collection location and the collection date and timeof the soil, or the physical properties of the collected sample,including the classification of the blood, the urine, the faeces, thebodily fluid, the cells, or the like, of the patient, the classificationof the food, the classification of the soil, or the classification ofthe polluted water for example. Examples of the information that managesthe samples include the collector and the collection date of the samplethereof, the contact person for the sample, and the inspection date ofthe sample thereof for example.

The “inspection information” represents information showing the contentof the inspection performed with respect to the sample, and can includeinspection items such as various genetic information (SNPs, basesequence determination for example), genetic testing, or other variousprotein information or the types of reagents utilized in the inspection,the production lot number of the reagents, the calibration curves forthe reagents, the type and the structure of the instruments for testing,or the type of biological material fixed to a carrier and the like, forexample. This information is displayed in a handwritten case, a printedcase, a case where it is a barcode, or a case where it is a QR(registered trademark) code (a matrix type two-dimensional code) forexample. The image data is analyzed, converted to analysis datacorresponding to the code data, and output.

A sixteenth aspect of the invention is a light measurement apparatus fora reaction vessel comprising: a nozzle head provided with asuction-discharge mechanism that performs suction and discharge ofgases, and one or two or more nozzles that detachably mount dispensingtips in which the suction and the discharge of liquids is possible bymeans of the suction-discharge mechanism; a vessel group having at thevery least one or two or more liquid housing parts that house reactionsolutions used for various reactions, a liquid housing part that housesa magnetic particle suspension in which magnetic particles that are ableto capture a target compound are suspended, a liquid housing part thathouses a sample, one or two or more liquid housing parts that house asolution for separating and extracting of the target compound, and twoor more reaction vessels; a nozzle head transfer mechanism that makes aninterval between the nozzle head and the vessel group relativelymovable; a magnetic force part that is able to adsorb the magneticparticles on an inner wall of dispensing tips mounted on the nozzles; alight guide stage provided to the nozzle head and having two or morelinking portions to which ends of one or two or more light guideportions, that are directly or indirectly linkable with the respectivereaction vessels and optically connect with the interior of the linkedreaction vessels, are provided; a connecting end arranging body havingan arranging surface that arranges and supports along a predeterminedpath two or more connecting ends, to which back ends of the light guideportions, in which front ends thereof are provided to the linkingportions, are provided, the connecting ends are provided correspondingto the respective linking portions; a measurement device having one ortwo or more measuring ends that are successively optically connectablewith the respective connecting ends along the predetermined path, thatis able to receive light based on an optical state within the reactionvessels by means of optical connections between the connecting ends andthe measuring ends, the measuring ends are provided approaching ormaking contact with the arranging surface; and a light guide switchingmechanism provided along the predetermined path of the connecting endarranging body that relatively moves the respective connecting ends andthe respective measuring ends such that they become successivelyoptically connected.

In a case where the measurement device comprises a plurality of types ofmeasurement devices, it is preferable to provide with respect to therespective measurement devices, and at the very least between adjacentmeasurement devices, a modulator that modulates the intensity of thelight to be received by the respective measurement devices at mutuallydifferent predetermined frequencies, and a tuner that, with regard tothe light received by the respective measurement devices, performstuning to the predetermined frequencies, and obtains the correspondingintensity of the received light.

Here, the “reaction solution” is for example an amplification solutionused for nucleic acid amplification. Furthermore, the “target compound”represents nucleic acids or the fragments thereof, which is theamplification subject. It is preferable to provide a tip detachingmechanism that detaches the sealing lids or the dispensing tips from thenozzles. In the present apparatus, it is preferable to provide a samplesupplying device having a dispensing function that supplies the samples,the reagents, the washing liquids, the buffers, and the like that arenecessary for the vessel group at a position separate to the stage ofthe light measurement apparatus for a reaction vessel, and to make thewhole stage, to which the supplied vessel group is built-in, beautomatically moved to the position of the stage of the lightmeasurement apparatus for a reaction vessel and to be made exchangeable.Consequently, processing, including preparation processing such as thedispensing processing or the supplying processing with respect to thevessel group, can be consistently performed.

It is possible to respectively combine the second aspect of theinvention through to the thirteenth aspect of the invention with thepresent aspect of the invention.

A seventeenth aspect of the invention is a light measurement method fora reaction vessel comprising: moving a linking portion provided with anend of one or two or more light guide portions, with respect to two ormore reaction vessels; simultaneously directly or indirectly linking thereaction vessels and the linking portions and optically connecting theinterior of the linked reaction vessels and the light guide portions;performing temperature control within the reaction vessels; guidinglight from the reaction vessels to a connecting end arranging bodyhaving an arranging surface that arranges and supports along apredetermined path two or more connecting ends, to which back ends ofthe light guide portions, in which front ends thereof are provided tothe linking portions, are provided, the connecting ends are providedcorresponding to the respective linking portions; and successivelyoptically connecting along the predetermined path the one or two or moremeasuring ends provided to a measurement device, which are providedapproaching or in contact with the arranging surface, and the respectiveconnecting ends, by moving the connecting end arranging body, to therebymake the measurement device receive light based on an optical statewithin the reaction vessels.

In a case where the measurement device comprises a plurality of types ofmeasurement devices, it is preferable, at the very least betweenadjacent measurement devices, to modulate the intensity of the light tobe received by the measurement devices at mutually differentpredetermined frequencies, and with regard to the light received by themeasurement devices, to perform tuning to the predetermined frequencies,and to obtain the corresponding intensity of the received light.

Furthermore, it is preferable for the front ends of a light guideportion bundle comprising a plurality of light guide portions to beprovided to the respective linking portions, the light guide portionbundle to be such that the rear ends of a portion of the light guideportion bundle are provided to first connecting ends of the connectingend arranging body, a portion or all of the remaining rear ends of thelight guide portion bundle to be provided to second ends of theconnecting end arranging body, the predetermined path to comprise afirst path and a second path, the connecting end arranging body to havea plurality of pairs of first connecting ends and second connecting endsthat are optically connected with the linking portion, and a single pairof a third connecting end and a fourth connecting end that is opticallymutually connected by means of the light guide portion, and at the timethe first measuring end provided to the measurement device relativelymoves along the first path, which comprises a plurality of firstconnecting ends and a single third connecting end, and the secondmeasuring end relatively moves along the second path, which comprisesthe second connecting ends and the fourth connecting end, for the firstmeasuring ends to optically connect with the irradiation source of themeasurement device, and the second measuring ends to connect with thelight receiving portion of the measurement device, and for each pair,for the first measuring end to successively connect with the pluralityof first connecting ends and the single third connecting end, and thesecond measuring end to successively connect with the plurality ofsecond connecting ends and the single fourth connecting end. The effectsdescribed above are also achieved in this case.

It is possible to respectively combine the second aspect of theinvention through to the fifteenth aspect of the invention with thepresent aspect of the invention.

An eighteenth aspect of the invention is a light measurement method fora reaction vessel, wherein the measurement device has a plurality oftypes of specific wavelength measurement devices that are able toreceive light of specific wavelengths or specific wavelength bands, andthe respective specific wavelength measurement devices have at least onemeasuring end that is successively optically connectable with theconnecting ends along the predetermined path. The method comprisesaligning measuring ends of a plurality of types by a measuring endaligning portion, and successively optically connecting the measuringends with the connecting ends along the path, to thereby make therespective specific wavelength measurement devices receive the light ofspecific wavelengths or specific wavelength bands based on an opticalstate within the reaction vessels.

A nineteenth aspect of the invention is a light measurement method for areaction vessel comprising simultaneously mounting two or more sealinglids, which have transparency, that are arranged in the vessel group andare finable with apertures of the reaction vessels, on reaction vessels,and then moving the light guide stage with respect to the sealing lidsof the reaction vessels.

A twentieth aspect of the invention is a light measurement method for areaction vessel comprising pressing or shaking with respect to thesealing lids covering the apertures of the reaction vessels.

A twenty-first aspect of the invention is a light measurement method fora reaction vessel comprising heating the sealing lids sealing thereaction vessels through the light guide stage.

A twenty-second aspect of the invention is a light measurement methodfor a reaction vessel comprising: directly or indirectly linkingapertures of the reaction vessels and the linking portions; and at thetime of performing temperature control within the reaction vessels,according to temperature control by a temperature controller, which hasa temperature source provided making contact with or approaching lowerside wall sections of the reaction vessels, heating upper side wallsections of the reaction vessels, which are positioned further on anupper side than the lower side wall sections, by means of a heat sourceof a heating portion, which is provided making contact with orapproaching the upper side wall sections, and thereby preventing director indirect condensation of the linking portions.

A twenty-third aspect of the invention is a light measurement method fora reaction vessel comprising: detachably mounting dispensing tips onrespective nozzles, which are provided to a nozzle head and perform thesuction and the discharge of gases; separating a target compound byusing a magnetic force part, a nozzle head transfer mechanism thatrelatively moves an interval between the nozzle head and a vessel group,a magnetic particle suspension housed within a vessel group, in whichmagnetic particles that are able to capture a target compound aresuspended, a sample, and a solution for separating and extracting of atarget compound; introducing the separated target compound and areaction solution used for reactions to a plurality of reaction vesselsprovided to a vessel group; moving a light guide stage, which has two ormore linking portions that are provided to the nozzle head and in whichfront ends of one or two or more light guide portions are also provided,with respect to apertures of the reaction vessels by means of, at thevery least, the nozzle head transfer mechanism; directly or indirectlysimultaneously linking the respective apertures of the reaction vesselsand the linking portions, and optically connecting the interior of thereaction vessels and the light guide portions that are linked;performing temperature control within the reaction vessels; guidinglight from the reaction vessels to a connecting end arranging body thatarranges and supports along a predetermined path two or more connectingends, to which back ends of the light guide portions, in which frontends thereof are provided to the linking portions, are provided, theconnecting ends are provided corresponding to the respective linkingportions; and successively optically connecting one or two or moremeasuring ends that are provided to a measurement device, and providedapproaching or making contact with an arranging surface, and theconnecting ends, along the predetermined path by relatively moving them,to thereby make the respective measurement devices receive the lightbased on an optical state within the reaction vessels.

It is possible to respectively combine the second aspect of theinvention through to the fifteenth aspect of the invention with thepresent aspect of the invention.

Furthermore, in a case where the measurement device comprises aplurality of types of measurement devices, it is preferable at the timethe light from the reaction vessels is guided to the connecting endarranging body, at the very least between adjacent measurement devices,for the intensity of the light to be received by the respectivemeasurement devices to be modulated at mutually different predeterminedfrequencies, and with regard to the light received by the respectivemeasurement devices, for tuning to be performed to the predeterminedfrequencies, and for the corresponding intensity of the received lightto be obtained.

According to the first aspect of the invention, the sixteenth aspect ofthe invention, the seventeenth aspect of the invention, or thetwenty-third aspect of the invention, as a result of linking with theplurality of reaction vessels by means of the linking portions providedto the light guide stage and optically connecting with the interior ofthe reaction vessels, the optical state within the reaction vessels istransmitted via the plurality of reaction vessels, the light guidestage, and the light guide portion, to the connecting ends of thearranging surface of the connecting end arranging body, and theconnecting ends arranged along the predetermined path on the arrangingsurface of the connecting end arranging body and the measuring ends ofthe measuring device are successively optically connected. Therefore,compared to a case where the measuring ends are directly scanned withrespect to the apertures of the reaction vessels, then in addition topreventing the attenuation or the leakage of light from the scatteringof light at the interval between the measuring ends and the liquidsurface, the arrangement of the connecting ends is such that it can berearranged in order to perform the connection with the measuring endsrapidly and smoothly, and with certainty. Therefore, measurements with ahigh reliability, and more efficient and rapid measurements of theoptical state within the reaction vessels, can be performed.

Consequently, with consideration of the stable light receivable time,the structure of the measuring ends, and the like, then the arrangingregion of the connecting ends as a whole, or the distance betweenadjacent connecting ends can be achieved by integration that makes thearranging region or the adjacent distances of the linking portionssmaller, and, in comparison to the arrangement of the linking portion,by the smoothing of the movement of the measuring ends as a result ofthe linearization or the expansion of the radius of curvature of thepredetermined path.

Switching of the optical system is performed by means of the movementbetween the measuring ends and the connecting ends on the arrangingsurface along the predetermined path. Therefore, the structure of theoptical system can be simplified. Furthermore, by separating theconnecting ends, the measuring ends, and the measurement device from thereaction vessels or the light guide stage, in which temperature controlor heating control is performed, thermal effects on the optical systemelements are excluded, and processing with a high reliability can beperformed.

The movement of the connecting ends with respect to the measuring endsincludes continuous or intermittent movement. As a result of themeasurement by real-time PCR, an amplification curve is created, whichcan be utilized in various analyses, such as the determination of theinitial concentration of DNA.

Moreover, since the measurement of the plurality of reaction vessels canbe performed in parallel with a single measuring device by utilizing thestable light receivable time, the expansion of the scale of theapparatus is suppressed by reducing the number of measuring devices, andthe manufacturing costs can be reduced. Further, since it is possible tomeasure, by successively moving the interval between the measuring endsand the connecting ends through the shortest distance along thepredetermined path determined beforehand, the measurements can beperformed in parallel by a simple mechanism of only a transfermechanism.

In a case where the reactions and the measurements are performed bysealing the reaction vessels by directly or indirectly linking theapertures of the reaction vessels with the linking portions, automaticmeasurements with a high reliability in which cross-contaminations andthe contamination of light can be prevented with certainty can beperformed.

Furthermore, according to the first aspect of the invention, thesixteenth aspect of the invention, the seventeenth aspect of theinvention, or the twenty-third aspect of the invention, in a case wherewith respect to a plurality of measurement devices, at the very leastbetween adjacent measurement devices, the intensity of the light to bereceived by the respective measurement devices is modulated at mutuallydifferent predetermined frequencies, and the light intensity isdemodulated from the received light, the measurement devices are suchthat optical cross-talk mainly due to the entry of excitation light fromadjacent measurement devices is prevented, and optical measurements witha high reliability can be performed. Consequently, since the arrangementof the connecting ends can be integrated even further, compact and rapidmeasurements can be performed.

According to the second aspect of the invention, at the time of movementwith respect to the connecting ends arranged on the connecting endarranging body and the measuring ends, the measurement device isimmovable with respect to the reaction vessels and the light guide stagethat is linked with the same. Therefore, at the time of a measurement, aload from an inertia force due to acceleration accompanying themovement, and the like, is not placed on the optical system elements orthe electronic elements built into the measurement device body,displacements of the optical system elements and destruction of theelectronic elements are prevented, and an accurate measurement with ahigh reliability can be performed. In cases other than a measurement,the measurement device body is movable with respect to the reactionvessels and the like. Therefore, it is possible to transport themeasurement device close to the reaction vessels and perform ameasurement.

According to the third aspect of the invention, in the case where thelight based on the optical state is a fluorescent light, the respectivespecific wavelength measurement devices are provided with irradiationsources of excitation light. Therefore, as the power source for theirradiation sources of the excitation light, an alternating currentvoltage of the predetermined frequency is applied, and excitation lightthat changes the intensity of light at the predetermined frequency canbe easily obtained. By so doing, fluorescent light having an intensitycorresponding to the irradiation of the excitation light, and thatchanges at the same frequency can be easily obtained. As a result, itbecomes possible to perform highly reliable measurement without crosstalk of light between adjacent measuring devices, with a simpleconfiguration.

According to the fourth aspect of the invention, the sixteenth aspect ofthe invention, or the twenty-third aspect of the invention, by providinga stage transfer mechanism that moves the light guide stage, it ispossible to simultaneously directly or indirectly link the linkingportions with the reaction vessels without human intervention.Therefore, cross-contamination is prevented, and processing can beefficiently performed.

According to the third aspect of the invention or the eighteenth aspectof the invention, by using a plurality of types of luminescentcompounds, colored compounds, color changing compounds, or lightvariation compounds, within a single reaction vessel, then for examplein a case where amplification processing is performed in parallel underthe same conditions on a plurality of types of amplification subjects,it is possible to perform multiplex PCR amplification or multiplexreal-time PCR on a plurality of types of amplification subjects by usinga primer labeled with a plurality of types of luminescent compounds andthe like. At that time, by combining the switching of the receiving ofthe light of a plurality of types of specific wavelengths or specificwavelength bands from the plurality of types of luminescent compoundsand the like, with a mechanism utilizing the stable light receivabletime that is used at the time of movement between the plurality ofreaction vessels, it is not necessary to separately provide a speciallight switching mechanism, and the apparatus mechanism can be simplifiedand manufacturing costs can be reduced. Furthermore, since therespective specific wavelength measuring devices each receive light of aspecific wavelength or a specific wavelength band, the effects of otherspecific wavelengths or specific wavelength bands are not received, andhigh-accuracy measurements can be performed. Moreover, since therespective specific wavelength measuring devices are each modularizedsuch that removal and addition can be performed, processing with a highversatility according to the processing aims can be performed.

According to the fifth aspect of the invention or the nineteenth aspectof the invention, by mounting the sealing lids arranged in the vesselgroup to the linking portions or the nozzles, it is possible to performmounting to the apertures of the reaction vessels by means of movementof the nozzle head and the like. Therefore the housed substances withinthe reaction vessels do not make direct contact with the linkingportions of the stage, and hence cross-contaminations can be effectivelyprevented. Furthermore, since it is not necessary to provide a dedicatedmechanism for mounting the sealing lids on the reaction vessels, thescale of the apparatus is not expanded, and the manufacturing costs arereduced.

According to the twentieth aspect of the invention, the sealing of thereaction vessels can be performed with certainty by controlling thesealing lids covering the apertures of the reaction vessels such thatthey are pressed. Furthermore, by shaking the sealing lids, the sealedstate between the apertures of the reaction vessels and the sealing lidscan be rapidly and easily removed and released. Therefore, a highprocessing efficiency and reliability can be obtained.

According to the sixth aspect of the invention or the twenty-firstaspect of the invention, by performing control such that the linkingportions are heated, condensation at the time of temperature control ofthe reaction vessels that are sealed by the sealing lids is prevented,and measurements via the sealing lids, which have transparency, can beperformed with certainty and a high accuracy.

According to the seventh aspect of the invention or the twenty-secondaspect of the invention, by performing heating of the upper side wallsections of the reaction vessels according to the temperature control ofthe lower side wall sections of the reaction vessels, the direct orindirect condensation of the linking portions can be prevented. In thiscase, the linking portions and the sealing lids are not directly heated,and heating is performed at the upper side wall sections of the reactionvessels. Therefore, the effects of direct heating toward the opticalsystem elements provided to the linking portions can be reduced orremoved. Consequently, in addition to reducing or removing imagedistortions and the like due to the degradation or the change inproperties of the optical system elements, various optical systemelements can be provided to the linking portions. Therefore, precisemeasurements with a high versatility can be performed. Furthermore, itis not necessary to provide a heating portion directly above thevessels, and the structure directly above the vessels, and therefore thestructure of the apparatus as a whole is simplified, and it is possibleto further approach the linking portions possessing optical systemelements, to the vessels and to perform the optical measurements withcertainty. With respect to the lower side wall sections, according tothe heating of the upper side wall sections, temperature control isperformed such that the temperatures are guided to the respectivepredetermined temperatures set using a coolable Peltier element and thelike, and measurements with a high reliability can be performed.

According to the eighth aspect of the invention, by using a material forthe linking portion that is usable to a temperature higher than the dewpoint of water vapor, has a low thermal conductivity, and a highrigidity, prevention of condensation is achieved. Furthermore, theeffects of heat toward the optical elements provided to the linkingportion are reduced, and optical processing with a high reliability canbe performed.

According to the ninth aspect of the invention, the thirteenth aspect ofthe invention, or the sixteenth aspect of the invention, as a result ofthe light guide stage being built into the nozzle head to which thenozzles are provided, a transfer mechanism (at the very least for the Xaxis and Y axis directions) between the reaction vessels of themeasuring device is not separately provided, and since it can becombined with the transfer mechanism of the nozzles, the expansion ofthe scale of the apparatus can be prevented. Furthermore, since thetransfer to the reaction vessel of the sample solution, the reagentsolutions, and the reaction solutions, which are to be housed within thereaction vessels, and which represent the measurement subject, and thepreparation, can be performed using the functions of the nozzles, stepsfrom the processing to the measurement of the measurement subject can beconsistently, efficiently, and rapidly performed.

According to the tenth aspect of the invention, since the sealing lidsare transferred by being mounted on the nozzles, a new, dedicated lidtransfer mechanism is not provided, and the expansion of the apparatusscale can be prevented by using existing mechanisms. On the other hand,in a case where the sealing lids are transferred or retained by beingmounted on the linking portions, it is possible to make the diameters ofthe linking portions and the nozzles different. Therefore, it becomespossible to provide various optical system elements within the linkingportions that are not limited by the size of the nozzles, and processingwith a high versatility and with reliability can be performed.

According to the eleventh aspect of the invention, by providing thefront end of a light guide portion bundle comprising a plurality oflight guide portions to the linking portions, dividing the light guideportion bundle into a plurality of bundles, and separating the back endof the light guide portions to a plurality of connecting ends, and bysimultaneously connecting with a plurality of measuring ends having oneor a plurality of measurement devices, the receiving of light of aplurality of types of wavelengths or wavelength bands, or theirradiation of excitation light with respect to the reaction vessels andthe receiving of light can be simultaneously performed. Therefore,processing of multiple fluorescent lights can be performed.

According to the twelfth aspect of the invention, the first measuringends optically connect with the irradiation source of the measurementdevice, and the second measuring ends optically connect with the lightreceiving portion of the measurement device, and in addition, the endsof the light guide portions that are connectable with the irradiationsource and the light receiving portion are mixed. Therefore, at the timeof a measurement of fluorescent light, it is possible to irradiateexcitation light within the reaction vessel without unevenness, and tomeasure the strength corresponding to the amount of fluorescence withcertainty.

According to the fourteenth aspect of the invention, by providingtraversable nozzles that are movable such that they traverse therespective exclusive regions, and dispensing target compounds or samplesof the same nucleic acids and the like with respect to a plurality ofexclusive regions, the same target compounds or samples can be utilizedin reactions in which the conditions are changed. Furthermore, bycombining the movement of the traversable nozzles with the transfermechanism of the connecting end arranging body, the expansion of theapparatus scale can be suppressed.

According to the fifteenth aspect of the invention, by displayinginformation at the respective exclusive regions, and together with themovement of the traversable nozzles, reading in the informationdisplayed at the respective exclusive regions with a camera, reactionand measurement processing with a high reliability can be performedwithout expanding the apparatus scale.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is an overall block-diagram showing a light measurement apparatusfor a reaction vessel according to a first embodiment of the presentinvention.

FIG. 2 is an overall perspective view showing the light measurementapparatus for a reaction vessel according to a first exemplaryembodiment.

FIG. 3 is a plan view showing enlarged, a vessel group of the lightmeasurement apparatus for a reaction vessel shown in FIG. 2.

FIG. 4 is a front view and a side view showing enlarged, a nozzle headof the light measurement apparatus for a reaction vessel shown in FIG.2.

FIG. 5 is a perspective view as viewed from the front side of the nozzlehead shown in FIG. 4.

FIG. 6 is a side view showing more specifically, the transfer mechanismsand the suction-discharge mechanism shown in FIG. 4.

FIG. 7 is a perspective view showing more specifically, thesuction-discharge mechanism 53 and the like shown in FIG. 4.

FIG. 8 is a perspective view as viewed from the rear side of the nozzlehead shown in FIG. 4.

FIG. 9 is a cross-sectional view showing a state in which the linkingportion shown in FIG. 4 is linked with a reaction vessel.

FIG. 10 is a drawing showing the specific wavelength measurement deviceshown in FIG. 4.

FIG. 11 is a perspective view showing a light guide stage, a connectingend arranging body, and a sealing lid transport mechanism of a lightmeasurement apparatus for a reaction vessel according to a secondexemplary embodiment.

FIG. 12 is an enlarged perspective view showing the light guide stageshown in FIG. 11 with a portion cut away.

FIG. 13 is an enlarged cross-sectional view of the linking portion shownin FIG. 12.

FIG. 14 is a cross-sectional view showing an example of the sealing lidshown in FIG. 11.

FIG. 15 is an enlarged perspective view of the sealing lid transportingbody shown in FIG. 11.

FIG. 16 is a cross-sectional view of the sealing lid transporting bodyshown in FIG. 15.

FIG. 17 is a perspective view as viewed from the lower side of thesealing lid transporting body shown in FIG. 15.

FIG. 18 is a schematic diagram showing an example of the positionalrelationship between the optical fiber ends provided on the linkingportions and the reaction vessel.

FIG. 19 is an overall block-diagram showing a light measurementapparatus for a reaction vessel according to a second embodiment of thepresent invention.

FIG. 20 is a side view showing more specifically, the transfer mechanismand the suction-discharge mechanism according to a first exemplaryembodiment of FIG. 19.

FIG. 21 is a cross-sectional view showing a state in which a linkingportion according to the first exemplary embodiment of FIG. 19 is linkedwith a reaction vessel.

FIG. 22 is a cross-sectional view showing a state in which a linkingportion according to a second exemplary embodiment of FIG. 19 is linkedwith a reaction vessel.

FIG. 23 is a cross-sectional view showing a state in which a linkingportion according to a third exemplary embodiment of FIG. 19 is linkedwith a reaction vessel.

FIG. 24 is a drawing showing heating control according to a fourthexemplary embodiment of FIG. 19.

FIGS. 25A, B and C are drawings showing the temperature-fluorescenceintensity characteristics within a reaction vessel according to a fifthexemplary embodiment of FIG. 19.

FIG. 26 is a drawing showing temperature measurement results of aferrule within a linking portion according to a sixth exemplaryembodiment of FIG. 19.

FIG. 27 is a block diagram showing a measurement device of a lightmeasurement apparatus for a reaction vessel according to a thirdembodiment of the present invention.

FIG. 28 is a drawing showing connection between the connecting endarranging body and a measuring end according to a first exemplaryembodiment of FIG. 27.

FIGS. 29A, B, C, D, and E are drawings showing the effects betweenmeasurement devices with regard to the light to be received by themeasurement devices of FIG. 27.

FIG. 30 is a drawing showing the spacing of the linking portions and thespacing of the connecting ends of the connecting end arranging body ofFIG. 27.

FIG. 31 is a drawing showing a measurement possibility of a measurementdevice for the case of FIG. 30.

FIG. 32 is a drawing showing connection between the connecting endarranging body and a measuring end according to a seventh embodiment ofthe present invention.

FIG. 33 is a graph showing fluorescence intensities within the reactionvessels using the connecting end arranging body according to FIG. 32,and the standardized fluorescence intensities.

FIG. 34 is a block diagram showing a measurement device of a lightmeasurement apparatus for a reaction vessel according to an eighthembodiment of the present invention.

BEST MODE FOR CARRYING OUT THE INVENTION

Next, an embodiment of the present invention is described with referenceto the drawings. This embodiment is not to be interpreted as limitingthe present invention unless particularly specified. Furthermore, in theembodiments or in the exemplary embodiments, the same objects aredenoted by the same reference symbols, and the descriptions are omitted.

FIG. 1 is shows a block-diagram of a light measurement apparatus for areaction vessel 10 according to a first embodiment of the presentinvention.

The light measurement apparatus for a reaction vessel 10 broadly has: avessel group 20 in which a plurality (twelve in this example) ofreaction vessel groups 23 _(i) (i=1, . . . , 12, omitted hereunder) arearranged; a nozzle head 50 that has a nozzle arranging portion 70 inwhich a plurality (twelve in this example) of nozzles 71 _(i) thatdetachably mount dispensing tips are arranged, and a light guide stage32 that has a plurality (twelve in this example) of linking portions 31_(i) provided with the ends of two or more light guide portions, whichhave a flexibility, that are directly or indirectly linkable with theapertures of the reaction vessels and optically connect to the linkedreaction vessel interior; a measurement device 40 that is provided fixedto the nozzle head 50; a nozzle head transfer mechanism 51 that makesthe nozzle head 50 movable in the X axis direction for example; atemperature controller 29 that performs predetermined temperaturecontrol with respect to the reaction vessel group 23 _(i) of the vesselgroup; a CPU+program 60 composed of a CPU, a ROM, a RAM, various typesof external memory, communication functions such as a LAN, and a programstored in the ROM, and the like; and a control panel 13 having a displayportion such as a liquid crystal display, and an operation portion, suchas operation keys or a touch panel.

The nozzle head 50 has: a stage Z axis transfer mechanism 35 that makesthe light guide stage 32 movable in the Z axis direction with respect tothe vessel group 20 independent of the nozzle arrangement portion 70; anozzle Z axis transfer mechanism 75 that makes the nozzle arrangementportion 70 movable in the Z axis direction with respect to the vesselgroup 20 independent of the stage 30; a magnetic force part 57 that, bymeans of a magnet 571 provided on a narrow diameter portion 211 ia of adispensing tip 211 i detachably mounted on the nozzle 71 i such that itcan approach and separate, is able to apply and remove a magnetic fieldwith respect to the interior; a suction-discharge mechanism 53 thatmakes the suction and the discharge of liquids with respect to thedispensing tip 211 i mounted on the nozzle 71 i possible by performingthe suction and the discharge of gases with respect to the nozzle 71 i;and a punching mechanism 55 which is driven by the suction-dischargemechanism 53, for punching a film that covers the apertures of theliquid housing parts of the vessel group 20 to seal various liquids inadvance. The stage transfer mechanism corresponds to the nozzle headtransfer mechanism and the stage Z axis transfer mechanism.

The nozzle head 50 further has a connecting end arranging body 30 thatarranges and supports a plurality (twelve in this example) of connectingends 34 _(i), which are provided corresponding to the respective linkingportions 31 _(i) and are provided with the back ends of optical fibers(bundle) 33 _(i), which represent light guide portions in which thefront ends thereof are provided to the linking portions 31 _(i), suchthat, as an arranging surface, they are integrated along a predeterminedpath (a linear path along the Y axis direction in this example) providedon a vertical plane at a narrower spacing than the spacing between thelinking portions 31 _(i). Furthermore, the connecting end arranging body30 is provided at a position that is separated from the light guidestage 32 and the reaction vessel group 23 _(i).

The measurement device 40 is able to respectively receive the light ofspecific wavelengths or specific wavelength bands of six types offluorescent light, and also has six types of specific wavelengthmeasurement devices 40 _(j) (j=1, . . . , 6, omitted hereunder) that areable to irradiate excitation light of six types of specific wavelengthsor specific wavelength bands that are irradiated for the emission oflight.

The respective specific wavelength measurement devices 40 _(j) havemeasuring ends 44 _(j) that are provided approaching or making contactwith the arrangement surface, and are successively connectable with therespective connecting ends 34 _(i) along the predetermined path (alinear path along the Y axis direction). Furthermore, the respectivemeasuring ends 44 _(j) have two ends, namely a first measuring end 42_(j) and a second measuring end 43 _(j) arranged along the Y axisdirection. The first measuring ends 42 _(j) optically connect with anirradiation source provided to the specific wavelength measurementdevices 40 _(j). The second measuring ends 43 _(j) optically connectwith a photoelectric device, such as a photomultiplier tube, provided tothe specific wavelength measurement devices 40 _(j).

Furthermore, the nozzle head 50 has an arranging body Y axis transfermechanism 41, which represents a light guide switching mechanism, thatmoves the connecting end arranging body 30 along the Y axis direction onthe nozzle head 50 such that the respective connecting ends 34 _(i)arranged on the connecting end arranging body 30 and the respectivemeasuring ends 44 _(j) are successively connected.

Moreover, the light guide stage 32 has a heater 37, which represents theheating portion, for preventing condensation of the ends of the linkingportions 31 _(i) or the mounted sealing lids 251 _(i), which havetransparency, by heating.

The vessel group 20 comprises a plurality (twelve in this example) ofexclusive regions 20 _(i), in which one (in this example, one groupcorresponds to one) nozzle enters and the other nozzles do not enter,that correspond to the respective nozzles. The respective exclusiveregions 20 _(i) have: a liquid housing part group 27 _(i) comprising aplurality of housing parts in which reagent solutions and the like arehoused or are housable; a sealing lid housing part 25 _(i) in which oneor two or more sealing lids 251 _(i), which have transparency, that aredetachably mounted on the nozzles are housed or are housable; andhousing parts for tips and the like 21 _(i) that house, a plurality ofdispensing tips 211 _(i) that are detachably mounted on the nozzles, andthe samples, and the like. The liquid housing part group 27 _(i) has, atthe very least, one or two or more liquid housing parts that house amagnetic particle suspension, and two or more liquid housing parts thathouse a solution for separating and extracting used for the separationand the extraction of nucleic acids or the fragments thereof Ifnecessary, it further has two or more liquid housing parts that house asolution for amplification used for the amplification of nucleic acids,and a liquid housing part that houses a sealing liquid for sealing thesolution for amplification housed in a PCR tube 231 _(i), whichrepresents the reaction vessel, within the PCR tube 231 _(i).

It is preferable for the exclusive regions 20 _(i) to display a barcodeas the sample information and the inspection information for identifyingthe exclusive regions 20 _(i). Furthermore, the nozzle head 50 isprovided with a single traversable nozzle 71 ₀ in which liquids aretransportable or dispensable by traversing (moving in the Y axisdirection) the exclusive regions 20 _(i), and suction and discharge ismade to be performed by a traversable nozzle suction-discharge mechanism17 that is separate from the suction-discharge mechanism 53.Consequently, the solution of DNA and the like housed in a givenexclusive region 20 _(i) can be dispensed or delivered to the otherexclusive regions 20 _(k) (k≠i). It is preferable for this movement inthe Y axis direction to be also used by the arranging body Y axistransfer mechanism 41.

The CPU+program 60 has, at the very least: a nucleic acid processingcontrol portion 63 that performs instructions for a series of processes,such as extraction and amplification with respect to nucleic acids orthe fragments thereof, and sealing of the solution for amplification,with respect to the temperature controller 29, the nozzle head transfermechanism 51, the tip detaching mechanism 59, the suction-dischargemechanism 53, the magnetic force part 57, and the nozzle Z axis transfermechanism 75; and a measurement control portion 61 that, after thenozzle head transfer mechanism 51 and the stage Z axis transfermechanism 35 are controlled such that the linking portions 31, aresimultaneously directly or indirectly linked with the apertures of theplurality (twelve in this example) of PCR tubes 231 _(i), instructs ameasurement by the measurement devices 40 _(j) by controlling thearranging body Y axis transfer mechanism 41 such that the optical fibers(bundle) 33 _(i), which represent the light guide portions of thelinking portions 31 _(i), and the first measuring ends 42 _(j) and thesecond measuring ends 43 _(j) of the measuring ends 44 _(j) of themeasurement devices 40 _(j) mentioned below are optically connected.

Furthermore, the nucleic acid processing control portion 63 has anextraction control part 65 and a sealing lid control part 67. Thenucleic acid processing control portion 63 has the extraction controlpart 65 that performs instructions with respect to the tip detachingmechanism 59, the suction-discharge mechanism 53, the magnetic forcepart 57, the nozzle Z axis transfer mechanism 75, the nozzle headtransfer mechanism 51, and the stage Z axis transfer mechanism 35, for aseries of processes with respect to the nucleic acids or the fragmentsthereof, and the sealing lid control part 67 that performs instructionswith respect to the stage Z axis transfer mechanism 35 and the nozzlehead transfer mechanism 51 for a sealing process by the sealing lids.

Hereunder, a more specific first embodiment of the light measurementapparatus for a reaction vessel 10 mentioned above according to anembodiment of the present invention, is described with reference to FIG.2 to FIG. 10. FIG. 2 is a see-through perspective view showing anexternal view of the light measurement apparatus for a reaction vessel10 according to the first embodiment of the present invention.

FIG. 2A is a drawing showing an external view of the light measurementapparatus for a reaction vessel 10, which has: an enclosure 11 with asize of 500 mm in depth (Y axis direction), 600 mm in width (X axisdirection), and 600 mm in height (Z axis direction) for example, inwhich the vessel group 20, the nozzle head a nozzle head transfermechanism 51 described in FIG. 1, and a CPU+program 60 are housed in theinterior; a control panel 13 provided on the enclosure 11; and a drawer15 to which a stage is provided.

FIG. 2B is a perspective view that sees through the interior of theenclosure 11, wherein the stage, into which the vessel group 20 isbuilt-in, is able to be drawn out to the exterior by means of the drawer15, and further, the nozzle head 50 is movably provided in the X axisdirection with respect to the vessel group 20 by means of the nozzlehead transfer mechanism 51 of FIG. 1.

FIG. 2B is a drawing showing that the nozzle head 50 is largely providedwith: various transfer mechanisms 52 having an arranging body Y axistransfer mechanism 41, a stage Z axis transfer mechanism 35, and anozzle Z axis transfer mechanism 75; a traversable nozzlesuction-discharge mechanism 17; the measuring device 40; a connectingend arranging body 30; an optical fiber (bundle) 33 _(i); and themagnetic force part 57. The traversable nozzle suction-dischargemechanism 17 and the traversable nozzles 71 ₀ are supported such thatthey are movable in the Y axis direction by means of the arranging bodyY axis transfer mechanism 41 such that they traverse the exclusiveregions 20 _(i).

FIG. 3 is a plan view showing enlarged, the vessel group 20 shown inFIG. 2. The vessel group 20 is one in which twelve exclusive regions 20i (i=1, . . . , 12), wherein the longitudinal direction thereof is alongthe X axis direction and housing parts are arranged in a single rowform, are arranged in parallel along the Y axis direction at a pitch of18 mm for example. The exclusive regions 20 _(i) are separately providedwith a cartridge vessel for PCR amplification 201 _(i), a cartridgevessel for nucleic acid extraction 202 _(i), and a cartridge vessel forhousing tips 203 _(i). The prevention of cross-contaminations betweenthe exclusive regions 20 _(i) is achieved by providing partition walls201 ₀, 202 ₀, and 203 ₀ on the cartridge vessels 201 _(i), 202 _(i), and203 _(i) of the exclusive regions 20 _(i) on the edge of one side alongthe X axis direction.

The cartridge vessel for PCR amplification 201 ₁ has: the PCR tubes 231₁, which represent the reaction vessel that are detachably linked withthe twelve linking portions 31 _(i) provided to the light guide stage32, via a single sealing lid 251 _(i) which has transparency; the liquidhousing parts 271 _(i) which house a buffer solution necessary for thePCR reaction; the sealing lid housing parts 25 _(i) which house thesealing lids 251 _(i); the housing part for tips and the like 21 _(i)that house the tips for punching for punching the film covering the PCRtubes 231 _(i) and the liquid housing parts 271 _(i), and the dispensingtips 211 _(i), and barcodes 81 _(i) that display the sample informationand the inspection information relating to the cartridge vessels for PCRamplification 201 _(i).

The cartridge vessels for nucleic acid extraction 202 _(i) has: sevenliquid housing parts 272 _(i) for example, that house various reagentsfor nucleic acid extraction; reaction vessels 232 _(i) that house theextracted nucleic acids; and barcodes 82 _(i) that display variousinformation, such as the sample information and the inspectioninformation, related to the cartridge vessel. The PCR tubes 231 _(i) andthe reaction vessels 232 _(i) are temperature controllable by means ofthe temperature controller 29.

The cartridge vessels for housing tips 203 _(i) has: a tip for punchingthat is able to punch the film covering the cartridge vessel for nucleicacid extraction 202 _(i); two small-quantity dispensing tips thatperform the dispensing of small quantities of liquids; housing parts fortips and the like 21 _(i) that house dispensing tips for separationsthat are able to perform separation by adsorbing magnetic particles onan inner wall by applying and removing a magnetic force from theexterior, and a barcode 83 _(i) that displays various informationrelating to the cartridge vessel 203 _(i).

The capacity of the PCR tube 231 _(i), which represents the reactionvessel, is of the order of approximately 200 μL, and the capacity of theother reaction vessels, liquid housing parts, and tubes is of the orderof approximately 2 mL.

The PCR tube 231 _(i) is used for the amplification of nucleic acids orthe fragments thereof, and temperature control is performed by means ofthe temperature controller 29 based on a predetermined amplificationmethod, such as a thermal cycle (from 4° C. to 95° C.) for example. ThePCR tube 231 _(i) is formed with two levels as shown in FIG. 9 forexample, and has a narrow-mouthed piping part 233 provided on the lowerside in which the solution for amplification 234 _(i) is housed, and awide-mouthed piping part 235 _(i) provided on the upper side in whichthe sealing lid 251 _(i) is fittable. The inner diameter of thewide-mouthed piping part 235 _(i) is 8 mm for example, and the innerdiameter of the aperture of the narrow-mouthed piping part 233 _(i) isapproximately 5 mm for example. The reaction vessels 232 _(i) housed inthe reaction tube housing holes are temperature controlled forincubation to a constant temperature of 55° C. for example.

The liquid housing part group 272 _(i) houses the solutions forseparating and extracting as follows. A first liquid housing part houses40 μL of Lysis 1, a second liquid housing part houses 200 μL of Lysis 2,a third liquid housing part houses 500 μL of a binding buffer solution,a fourth liquid housing part houses a magnetic particle suspension, afifth liquid housing part houses 700 μL of a washing liquid 1, a sixthliquid housing part houses 700 μL of a washing liquid 2, a seventhliquid housing part houses 50 μL of distilled water as a dissociationliquid, and an eighth liquid housing part, which is slightly separated,houses 1300 μL of isopropyl alcohol (isopropanol) used for the removalof protein and the like, as a portion of the solution for separating andextracting protein. The respective reagents and the like are prepackedas a result of the punchable film covering the respective aperturesthereof.

In addition, 1.2 mL of distilled water is housed in a separate distilledwater reservoir, and tubes that house suspensions of bacteria, cells,and the like, or samples such as whole blood, are separately preparedfor each of the respective exclusive regions 20 _(i).

FIG. 4 is a front view and a side view of the nozzle head 50 accordingto the first embodiment of the present invention, and FIG. 5 is aperspective view from the front side.

The nozzle head 50 is one having: a nozzle arranging portion 70 in whichtwelve nozzles 71 _(i) are arranged; a tip detaching mechanism 59 thatis able to detach dispensing tips 211 _(i) mounted on the nozzles 71_(i); a suction-discharge mechanism 53; a magnetic force part 57 havingtwelve magnets 571 provided such that they are able to approach andseparate with respect to the dispensing tips 211 _(i); a light guidestage 32; twelve linking portions 31 _(i) provided to the light guidestage 32; a transfer mechanism portion 52 having a nozzle Z axistransfer mechanism 75 and a stage Z axis transfer mechanism 35; opticalfibers (bundles) 33 _(i) representing flexible light guide portions thatextend to the rear side from the linking portions 31 _(i); a connectingend arranging body 30; the arranging body Y axis transfer mechanism 41;a measuring device 40 having a measuring end 44; a traversable nozzle 71₀; and a suction-discharge mechanism 17 thereof.

The nozzle arranging portion 70 is provided with a cylinder supportingmember 73 that supports twelve cylinders 531 _(i) such that they arearranged along the Y axis direction at a predetermined pitch of 18 mmfor example. The nozzles 71 _(i) are provided on the downward end of thecylinders 531 _(i) such that they are communicated with the cylinders531 _(i).

The tip detaching mechanism 59 is provided with detaching shafts 593 onboth sides, and has a tip detaching member 591 that detaches the twelvedispensing tips 211 _(i) from the nozzles 71 _(i) by sliding in thevertical direction.

As shown specifically in FIG. 6 and FIG. 7, the tip detaching member 591is interlocked with the lowering of two tip detaching shafts 593 anddetaches the dispensing tips 211 _(i) from the nozzles 71 _(i). The tipdetaching shaft 593 is elastically supported by the cylinder supportmember 73 by means of a spring 600 wrapped around the outer peripherysuch that it is biased in the upward direction, and the upper endthereof is positioned above the upper ends of the cylinders 531 _(i) butbelow the lower limit position of the vertical movement range of thenormal suction and discharge of a cylinder drive plate 536 mentionedbelow. The two tip detaching shafts 593 are pushed in the downwarddirection by means of the cylinder drive plate 536 exceeding thevertical movement range and being lowered near the upper end of thecylinder 531 _(i), thus lowering the tip detaching member 591. The tipdetaching member 591 has twelve holes having an inner diameter that islarger than the outer diameter of the nozzles 71 _(i) but smaller thanthe mounting portions 211 _(i) c, which represents the largest outerdiameter of the dispensing tips 211 _(i), arranged at the pitchmentioned above such that the nozzles 71 _(i) pass therethrough.

As shown specifically in FIG. 6 and FIG. 7, the suction-dischargemechanism 53 has: the cylinders 531 _(i) for performing suction anddischarge of gases with respect to the dispensing tips 211 _(i) whichare communicated with the nozzles 71 _(i) and mounted on the nozzles 71_(i), and a piston rod 532 that slides within the cylinders 531 _(i); adrive plate 536 that drives the piston rod 532; a ball screw 533 thatthreads with the drive plate 536; a nozzle Z axis movable body 535 that,in addition to axially supporting the ball screw 533, is integrallyformed with the cylinder support member 73; and a motor 534 mounted onthe nozzle Z axis movable body 535 that rotatingly drives the ball screw533.

The magnetic force part 57 has a magnet 571 that is provided such thatit can approach and separate with respect to the narrow diameterportions 211 _(i) a of the dispensing tips 211 _(i) detachably mountedon the nozzles 71 _(i), and is able to apply and remove a magnetic fieldin the interior of the dispensing tips 211 _(i).

As shown specifically in FIG. 6, the nozzle Z axis transfer mechanism 75has: a ball screw 752 that threads with the Z axis movable body 535 andvertically moves the Z axis movable body 535 along the Z direction; anozzle head substrate 753 that axially supports the ball screw 752, andin addition to axially supporting the magnet 571 on the lower sidethereof such that it is movable in the X axis direction, is itselfmovable in the X axis direction by means of the nozzle head transfermechanism 51 mentioned below; and a motor 751 provided on the upper sideof the nozzle head substrate 753 that rotatingly drives the ball screw752.

As shown specifically in FIG. 6, the light guide stage 32 comprises ahorizontal plate 32 a and a vertical plate 32 b, which are letter-Lshaped plates in cross-section, and is provided with twelvecylinder-shaped linking portions 31 _(i) having front ends of opticalfibers (bundles) 33 _(i), which are directly or indirectly linkable withthe apertures of the PCR tubes 231 _(i) and are optically connected withthe interior of the linked PCR tubes 231 _(i), protruding in thedownward direction from the horizontal plate 32 a. Furthermore, a heater37 that heats the sealing lids 251 _(i) mounted on the linking portions31 _(i) and prevents condensation, is built into the bases of thelinking portions 31 _(i). The temperature of the heater is set toapproximately 105° C. for example. Since the light guide stage 32 issupported by the nozzle head substrate 753 by means of the nozzle headstage Z axis transfer mechanism 35 such that it is movable in the Z axisdirection, it is movable in the nozzle X axis direction and Z axisdirection.

The stage Z axis transfer mechanism 35 has: a side plate 355 provided onthe nozzle head substrate 753; a mount driving band-shaped member 354that is supported by a timing belt 352 spanning between two sprockets353 arranged in the vertical direction axially supported by the sideplate 355, and vertically moves in the Z axis direction; and a motorattached to the rear side of the nozzle head substrate 753 thatrotatingly drives the sprockets 353.

As shown in FIG. 7, the traversable nozzle suction-discharge mechanism17 is provided with a tip detaching mechanism 592 on the lower side ofthe suction-discharge mechanism 17 and on the upper side of the nozzle71 ₀. Furthermore, the suction-discharge mechanism 17 is provided with adigital camera 19. The suction-discharge mechanism 17 is movablyprovided in the Y axis direction by being attached to a timing belt 171spanning between two sprockets 173 that are rotatingly driven by a motor172.

FIG. 8 represents two perspective views of the nozzle head according tothe first embodiment viewed from the rear side, which show theconnection starting position (FIG. 8A) and the connection finishingposition (FIG. 8B) at the time the respective connecting ends of theconnecting end arranging body 30 and the respective measuring ends aresuccessively optically connected.

There are provided a connecting end arranging body 30 in which theconnecting ends 34, provided corresponding to the respective linkingportions 31, to which the front ends of the optical fibers (bundles) 33_(i) which pass through the horizontal plate 32 a of the light guidestage 32, are provided, and provided with the back ends thereof, arearranged on an arranging surface on a path along a straight line in theY axis direction, which represents a predetermined path, at a shorterspacing than the spacing of the linking portions 31 _(i); and sixmeasuring ends that are provided in the vicinity of, or making contactwith, the arranging surface, and are successively optically connectablewith the connecting ends 34 _(i) along the straight line. There is alsoprovided a measuring device 40 in which, by means of optical connectionsbetween the connecting ends and the measuring ends, the fluorescentlight within the PCR tubes 231 _(i), which represents the optical state,is receivable, and excitation light is also able to be irradiated.

Furthermore, the light guide stage 32 has a cylinder-shaped body 311_(i), which retains the optical fibers (bundles) 33 _(i) extending tothe rear side from the linking portions 31 _(i) such that they passthrough the interior in order to prevent folding, protrudingly providedupward from the horizontal plate 32 a directly above the linkingportions 31 _(i). In the same manner, the connecting end arranging body30 is also provided with a cylinder-shaped body 301 _(i), which retainsthe optical fibers (bundles) 33 _(i) extending from the connecting ends34 _(i) such that they pass through the interior in order to preventfolding, on the connecting end 34 _(i) side.

The arranging body Y axis transfer mechanism 41 that moves theconnecting end arranging body 30 in the Y axis direction has: arms 412and 413 provided to the connecting end arranging body 30; a joining body411 that joins the arms 412 and 413 and the timing belt; a guide rail414 that guides the Y axis movement of the joining body 411; and twosprockets spanned by the timing belt and arranged along the Y axisdirection.

The measuring device 40 is one that supports the measurement offluorescent light and comprises six types of specific wavelengthmeasuring devices 40 _(j) that are linearly aligned along a straightline in the Y axis direction, which represents the predetermined path,such that they support the measurement of six types of fluorescentlight, and they are provided fixed on a substrate of the nozzle head 50,such as the frame that encloses the transfer mechanism portion 52, or amember that supports the same. Therefore, depending on the mechanismprovided to the transfer mechanism portion 52, the measuring device 40does not move.

The measuring device 40 is one in which the measuring ends of theplurality of types (six in this example) of specific wavelengthmeasuring devices 40 _(j) (j=1, 2, 3, 4, 5, 6), and therefore, in thiscase, the specific wavelength measuring devices 40 _(j) themselves arealigned in a single row form, and integrally fixed to a member joinedwith the nozzle head substrate 753 using fixtures 45 _(j). The specificwavelength measuring devices 40 _(j) have: measuring ends 44 _(j)arranged along a straight line path in the Y axis direction whichrepresents the predetermined path, such that they successively opticallyconnect to the connecting ends 34 _(i); light detectors 46 _(j) in whichan optical system component having an irradiation source that irradiatesexcitation light to the PCR tubes 231 _(i) and a light receiving portionthat receives the fluorescent light generated in the PCR tubes 231 _(i)are built-in; and circuit boards 47 _(j). The measuring ends 44 _(j)have first measuring ends 42 _(j) that optically connect with theirradiation source, and second measuring ends 43 _(j) that opticallyconnect with the light receiving portion. Here, the light detectors 46_(j) and the circuit boards 47 _(j) correspond to the measuring devicemain body.

The pitch between the respective connecting ends 34 _(j), assuming apitch between the linking portions 31 _(i) of 18 mm, is 9 mm, which ishalf thereof. Then, the pitch between the measuring ends 44 _(j) is 9 mmor less for example.

There is a case where the first measuring ends 42 _(j) and the secondmeasuring ends 43 _(j) of the measuring ends 44 _(j) of the respectivespecific wavelength measuring devices 40 j are arranged aligned in alateral direction (Y axis direction) along the straight line of the Yaxis direction along the predetermined path, and a case where they arearranged aligned in a longitudinal direction (X axis direction). In theformer case, without stopping the emission of the excitation light, therespective measuring devices successively receive light at a timing forreceiving light determined based on the speed of the connecting endarranging body, the pitch between the connecting ends, the distancebetween the first measuring ends and the second measuring ends of themeasuring ends, and the pitch between the measuring ends.

On the other hand, in the latter case, as shown in FIG. 8, with respectto the connecting end, a first connecting end and a second connectingend are provided. The first connecting end connects only with the firstmeasuring ends 42 _(j), and the second measuring ends 43 _(j) connectonly with the second connecting end. The fixed path represents twopaths. Furthermore, the optical fibers (bundles) 33 _(i) have opticalfibers (bundles) 331 _(i) for receiving light that have the firstconnecting end, and optical fibers (bundles) 332 _(i) for irradiationthat have the second connecting end. In this case, compared to theformer case, connection with the linking portions is performed by meansof optical fibers in which the irradiation source and the lightreceiving portion are dedicated, and therefore, the control is simple,and the reliability is high since optical fibers that are respectivelysuitable for irradiation and receiving light can be used.

The speed of the connecting end arranging body 30 with respect to themeasuring ends 44 _(j) is determined with consideration of the stablelight receivable time, the lifetime of the fluorescent light withrespect to excitation light irradiation, the number of connecting ends,the pitch between the connecting ends, and the like (the distance of thepredetermined path). In the case of a real-time PCR measurement, it iscontrolled such that it becomes 100 mm to 500 mm per second for example.In the present embodiment, since the movement is performed by slidingthe arranging surface with respect to the measuring ends 44, theincidence of optical noise to the measuring ends 44 can be prevented.Furthermore, the connecting end arranging body 30 moves with respect tothe measuring ends intermittently such that it momentarily stops at eachpitch advance between the connecting ends or between the measuring ends,or continuously.

FIG. 9A is a drawing showing a state in which the linking portion 31 i(here, i=1 for example) that protrudes on the lower side from thehorizontal plate 32 a of the light guide stage 32 is indirectly linkedwith the PCR tube 231 i via the sealing lid 251 i, which hastransparency, that is mounted on the aperture of the PCR tube 231 i inthe exclusive region 20 i, and the linking portion 31 i is insertedwithin the indentation of the sealing lid 251 i, and the end surfacethereof is adhered to the bottom surface of the indentation of thesealing lid 251 i. The PCR tube 231 i comprises a wide-mouthed pipingpart 235 i and a narrow-mouthed piping part 233 i that is communicatedwith the wide-mouthed piping part 235 i and is formed narrower than thewide-mouthed piping part 235 i. Furthermore, the narrow-mouthed pipingpart 233 i is dried beforehand, or a liquid state solution foramplification 234 i is housed beforehand. Here, the reagent forreal-time amplification represents 70 μL, of a master mix (SYBR(registered trademark) Green Mix) consisting of enzymes, buffers,primers, and the like.

For the aperture of the wide-mouthed piping part 235 i, since thesealing lid 251 i that protrudes on the lower side of the sealing lid251 i, which has transparency, is mounted on the reaction vessel, it ismounted to the reaction vessel as a result of a tubular sealing portion252 i, which encloses the center portion of the sealing lid 251 i inwhich light passes through, being fitted. At the time the sealingportion 252 i is fitted, it is preferable for the diameter of theoptical fibers (bundle) 33 i, which represents the light guide portionthat passes through the interior of the linking portion 31 i, to be thesame or larger than the diameter of the aperture of the narrow-mouthedpiping part 233 i. Consequently, it becomes possible to receive thelight from the PCR tube 231 i with certainty. The narrow-mouthed pipingpart 233 i is housed within a block for temperature control that isheated or cooled by means of the temperature controller 29.

In this example, the optical fibers (bundle) 33 i comprise opticalfibers (bundle) for irradiation 332 i that are connectable with thesecond measuring end 43 j and optical fibers (bundle) for receivinglight 331 i that are connectable with the first measuring end 42 j.

FIG. 9B is a drawing showing an example in which the optical fibers(bundle) 33 i comprise an optical fiber bundle in which an optical fiberbundle comprising a plurality of optical fibers for receiving light thatare connectable with the second measuring end 43 j, and an optical fiberbundle comprising a plurality of optical fibers for irradiation that areconnectable with the first measuring end 42 j, are combined such thatthey become uniform.

It is preferable for a feeding device for samples and the like to beintegrated to the light measurement apparatus for a reaction vessel 10.The feeding device for samples and the like is a device for dispensingand supplying parent samples and the like with respect to the vesselgroup 20, and the stage in which is integrated the vessel group 20 towhich the parent samples and the like have been supplied, isautomatically moved to the light measurement apparatus for a reactionvessel and the like. The feeding device for samples and the like, forexample, has a parent vessel group that houses parent samples and thelike, a tip detaching mechanism, a suction-discharge mechanism, and asingle nozzle that, in addition to the suction and the discharge ofgases being performed by means of the mechanisms, is detachably mountedwith dispensing tips 211 i. Furthermore, it has a nozzle head providedwith a mechanism that moves along the Z axis direction with respect tothe parent vessel group and the housing part group for tips and the like21 of the vessel group 20, an X axis movable body provided with a Y axistransfer mechanism that moves the nozzle head in the Y axis directionwith respect to the parent vessel group and the like, an X axis transfermechanism that moves the X axis movable body along the X axis withrespect to the parent vessel group and the like, and the parent vesselgroup. It is preferable for the parent vessel group to have a parentsample housing part group arranged in a 12 row×8 column matrix form thathouses the parent samples to be supplied to the housing part group fortips and the like 21 of the vessel group 20, a distilled water andwashing liquid group, and a reagent bottle group.

FIG. 10 is a drawing showing a light detector 46 ₁ of a single specificwavelength measurement device 40 ₁ belonging to the measurement device40 according to a first exemplary embodiment of the present invention.

The specific wavelength measurement device 40 ₁ according to the presentexemplary embodiment, in addition to having an optical fiber 469 forexcitation light to be outgoing to the PCR tube 231 i and an opticalfiber 479 for light from the PCR tube 231 i to be incoming, has ameasuring end 441 provided on the lower ends of a first measuring end421 of the optical fiber 469 and a second measuring end 431 of theoptical fiber 479, an irradiation portion 462 that has a LED 467 thatirradiates excitation light through the optical fiber 469 and a filter468, and a light receiving portion that has an optical fiber 479, a drumlens 478, a filter 477, and a photodiode 472. This example shows a casewhere the first measuring end 421 and the second measuring end 431 areprovided along a direction (X axis direction) perpendicular to thestraight line of the Y axis direction, which represents thepredetermined path.

Next, a series of processing operations that perform real-time PCR ofthe nucleic acids of a sample containing bacteria using the lightmeasurement apparatus for a reaction vessel 10 according to theembodiment is described. Step S1 to step S13 below correspond toseparation and extraction processing.

In step S1, the drawer 15 of the light measurement apparatus for areaction vessel 10 shown in FIG. 2 is opened, the vessel group 20 ispulled out, and by utilizing a feeding device for samples and the like,which is provided separately from the vessel group 20, the samples whichare subject to testing, various washing liquids, and various reagents,are supplied beforehand, and furthermore, a liquid housing part in whichreagents and the like are prepacked is mounted.

In step S2, following returning of the vessel group 20 and closing ofthe drawer 15, the start of the separation and extraction andamplification processing is instructed by means of the operation of thetouch panel of the control panel 13 for example.

In step S3, the extraction control part 65 provided to the nucleic acidprocessing controller 63 of the CPU+program 60 of the light measurementapparatus for a reaction vessel 10 instructs the nozzle head transfermechanism 51 and moves the nozzle head 50 in the X axis direction,positions the tip for punching mounted to the nozzle 71 _(i) above thefirst liquid housing part of the liquid housing part group 27 _(i) ofthe vessel group, and punches the film covering the aperture of theliquid housing part by lowering the nozzle by means of the nozzle Z axistransfer mechanism 75, and in the same manner, the other liquid housingparts of the liquid housing part group 27 _(i) and the reaction vesselgroup 23 _(i) are successively punched by moving the nozzle head 50 inthe X axis direction.

In step S4, the nozzle head 50 is again moved in the X axis directionand moved to the housing part for tips and the like 21 _(i), and thenozzles 71 _(i) are lowered by means of the nozzle Z axis transfermechanism 75, and the dispensing tips 211 _(i) are mounted. Next, afterbeing raised by the nozzle Z axis transfer mechanism 75, the dispensingtips 211 _(i) are moved along the X axis by means of the nozzle headtransfer mechanism 51, and advanced to the eighth liquid housing part ofthe liquid housing part group 27 _(i). Then a predetermined amount ofisopropanol is aspirated from the liquid housing part, and they areagain moved along the X axis direction, and predetermined amounts arerespectively dispensed into the solution components (NaCl, SDSsolutions) housed in the third liquid housing part and the fifth liquidhousing part, and the distilled water housed in the sixth liquid housingpart, so that 500 μL of a binding buffer solution (NaCl, SDS,isopropanol), 700 μL of a washing liquid 1 (NaCl, SDS, isopropanol), and700 μL of a washing liquid 2 (water 50%, isopropanol 50%) arerespectively prepared as solutions for separating and extracting withinthe third, the fifth, and the sixth liquid housing parts.

In step S5, following movement to, among the housing parts for tips andthe like 21 _(i), the sample tube in which the sample is separatelyhoused, the narrow diameter portion 211 _(i) a of the dispensing tip 211_(i) is loweringly inserted using the nozzle Z axis transfer mechanism75, and, with respect to the suspension of the sample housed in thesample tube, following suspension of the sample within the liquid byrepeating the suction and the discharge by raising and lowering thedrive plate 536 of the suction-discharge mechanism 53, the samplesuspension is aspirated within the dispensing tip 211 _(i). The samplesuspension is moved along the X axis by means of the nozzle headtransfer mechanism 51 to the first liquid housing part of the liquidhousing part group 27 _(i) housing the Lysis 1 (enzyme) representing thesolution for separating and extracting, and the narrow diameter portion211 _(i) a of the dispensing tip 211 _(i) is inserted through the holein the punched film, and the suction and the discharge is repeated inorder to stir the sample suspension and the Lysis 1.

In step S6, the entire amount of the stirred liquid is aspirated by thedispensing tip 211 _(i), and incubation is performed by housing it inthe reaction vessel 232 _(i) comprising the reaction tubes retained inthe housing holes, that is set to 55° C. by means of the constanttemperature controller. Consequently, the protein contained in thesample is broken down and made a low molecular weight. After apredetermined time has elapsed, the reaction mixture is left in thereaction tube, the dispensing tip 211 _(i) is moved to the second liquidhousing part of the liquid housing part group 27 _(i) by means of thenozzle head transfer mechanism 51, and the entire amount of the liquidhoused within the second liquid housing part is aspirated by using thenozzle Z axis transfer mechanism 75 and the suction-discharge mechanism53, and it is transferred using the dispensing tip 211 _(i) by means ofthe nozzle head transfer mechanism 51, and the reaction solution isdischarged within the third liquid housing part by penetrating the holein the film and inserting the narrow diameter portion.

In step S7, the binding buffer solution housed within the third liquidhousing part, which represents a separation and extraction solution, andthe reaction solution are stirred, the solubilized protein is furtherdehydrated, and the nucleic acids or the fragments thereof are dispersedwithin the solution.

In step S8, using the dispensing tip 211 _(i), the narrow diameterportion thereof is inserted into the third liquid housing part bypassing through the hole in the film, the entire amount is aspirated andthe dispensing tip 211 _(i) is raised by means of the nozzle Z axistransfer mechanism 75, and the reaction solution is transferred to thefourth liquid housing part, and the magnetic particle suspension housedwithin the fourth liquid housing part is stirred with the reactionsolution. A cation structure in which Na+ ions bind to the hydroxylgroups formed on the surface of the magnetic particles contained withinthe magnetic particle suspension is formed. Consequently, the negativelycharged DNA is captured by the magnetic particles.

In step S9, the magnetic particles are adsorbed on the inner wall of thenarrow diameter portion 211 _(i) a of the dispensing tip 211 _(i) byapproaching the magnet 571 of the magnetic force part 57 to the narrowdiameter portion 211 _(i) a of the dispensing tip 211 _(i). In a statein which the magnetic particles are adsorbed on the inner wall of thenarrow diameter portion 211 _(i) a of the dispensing tip 211 _(i), thedispensing tip 211 _(i) is raised by means of the nozzle Z axis transfermechanism 75 and moved from the fourth liquid housing part to the fifthliquid housing part using the nozzle head transfer mechanism 51, and thenarrow diameter portion 211 _(i) a is inserted by passing through thehole in the film.

In a state in which the magnetic force within the narrow diameterportion 211 _(i) a is removed by separating the magnet 571 of themagnetic force part 57 from the narrow diameter portion 211 _(i) a ofthe dispensing tip 211 _(i), by repeating the suction and the dischargeof the washing liquid 1 (NaCl, SDS, isopropanol) housed in the fifthliquid housing part, the magnetic particles are released from the innerwall, and the protein is washed by stirring within the washing liquid 1.Thereafter, in a state in which the magnetic particles are adsorbed onthe inner wall of the narrow diameter portion 211 _(i) a as a result ofapproaching the magnet 571 of the magnetic force part 57 to the narrowdiameter portion 211 _(i) a of the narrow diameter portion 211 _(i) aagain, the dispensing tip 211 _(i) is, by means of the nozzle Z axistransfer mechanism 75, moved from the fifth liquid housing part to thesixth liquid housing part by means of the nozzle head transfer mechanism51.

In step S10, the narrow diameter portion 211 _(i) a of the dispensingtip 211 _(i) is inserted by passing through the hole in the film usingthe nozzle Z axis transfer mechanism 75. By repeating the suction andthe discharge of the washing liquid 2 (isopropanol) housed in the sixthliquid housing part in a state in which the magnetic force within thenarrow diameter portion 211 _(i) a is removed by separating the magnet571 of the magnetic force part 57 from the narrow diameter portion 211_(i) a of the dispensing tip 211 _(i), the magnetic particles arestirred within the liquid, the NaCl and the SDS is removed, and theprotein is washed. Thereafter, in a state in which the magneticparticles are adsorbed on the inner wall of the narrow diameter portion211 _(i) a by approaching the magnet 571 of the magnetic force part 57to the narrow diameter portion 211 _(i) a of the dispensing tip 211 _(i)again, the dispensing tip 211 _(i) is, following raising by means of thenozzle Z axis transfer mechanism 75, moved from the sixth liquid housingpart to the seventh liquid housing part in which the distilled water ishoused, by means of the nozzle head transfer mechanism 51.

In step S11, the narrow diameter portion 211 _(i) a of the dispensingtip 211 _(i) is lowered through the hole by means of the nozzle Z axistransfer mechanism 75, and by repeating the suction and the discharge ofthe distilled water at a slow flow rate in a state where the magneticforce is applied within the narrow diameter portion 211 _(i) a of thedispensing tip 211 _(i), the washing liquid 2 (isopropanol) issubstituted by water and is removed. Thereafter, by stirring themagnetic particles by repeating the suction and the discharge within thedistilled water which represents the dissociation liquid, in a state inwhich the magnet 571 of the magnetic force part 57 is separated from thenarrow diameter portion 211 _(i) a of the dispensing tip 211 _(i) andthe magnetic force is removed, the nucleic acids or the fragmentsthereof retained by the magnetic particles are dissociated (eluted) fromthe magnetic particles into the liquid. Thereafter, a magnetic field isapplied within the narrow diameter portion and the magnetic particlesare adsorbed on the inner wall by approaching the magnet 571 to thenarrow diameter portion 211 _(i) a of the dispensing tip 211 _(i), andthe solution containing the extracted nucleic acids, and the like, ismade to remain in the eighth liquid housing part. The dispensing tip 211_(i) is moved to the storage part of the housing parts for tips and thelike 21 _(i) in which the dispensing tip 211 _(i) was housed, by meansof the nozzle head transfer mechanism 51, and the dispensing tip 211_(i) to which magnetic particles are adsorbed, is detached from thenozzle 71 _(i) together with the magnetic particles and dropped into thestorage part, using the detaching member 591 of the tip detachingmechanism 59.

The following step S12 to step S15 corresponds to nucleic acidamplification and measurement processing.

In step S12, a new dispensing tip 211 _(i) is mounted on the nozzle 71_(i), the solution housed within the eighth liquid housing part, whichcontains nucleic acids and the like, is aspirated, and by transferringit to the PCR tube 231 _(i) in which the solution for amplification 234_(i) is housed beforehand, and discharging it, it is introduced into thevessel. As a result of moving the nozzle head 50 by means of the nozzlehead transfer mechanism 51, the nozzle 71 _(i) is moved above thesealing lid housing part 25 _(i) of the vessel group 20, which housesthe sealing lid 251 _(i). Mounting is performed by lowering using thenozzle Z axis transfer mechanism 75 and fitting an indentation forlinking 258 _(i) on the upper side of the sealing lid 251 to the lowerend of the nozzle 71 _(i). After being raised by the nozzle Z axistransfer mechanism 75, the sealing lid 251 is positioned above the PCRtube 231 _(i) using the nozzle head transfer mechanism 51, and bylowering the sealing lid 234 _(i) by means of the nozzle Z axis transfermechanism 75, it is fitted with the aperture of the wide-mouthed pipingpart 235 _(i) of the PCR tube 231 _(i), mountingly sealing it.

In step S13, the nozzle head transfer mechanism 51 is instructed bymeans of an instruction from the measurement control portion 61, and bymoving the nozzle head 50 along the X axis, the linking portion 31 _(i)of the light guide stage 32 is positioned above the PCR tube 231 _(i),which is mounted with the sealing lid 251 _(i). Then, by lowering thelight guide stage 32 by means of the stage Z axis transfer mechanism 35,the linking portion 31 _(i) is inserted into the indentation of thesealing lid 251 _(i), and the lower end thereof is made to make contactwith, or adhere to, the bottom surface of the indentation.

In step S14, due to an instruction by the nucleic acid processingcontroller 63, the temperature controller 29 instructs a temperaturecontrol cycle by real-time PCR, such as a cycle in which the PCR tube231 _(i) is heated for five seconds at 96° C. and heated for 15 secondsat 60° C., to be repeated forty nine times for example.

In step S15, when temperature control at each cycle by the nucleic acidprocessing controller 63 is started, the measurement control portion 61determines the start of elongation reaction processing at each cycle,and instructs the continuous or intermittent movement of the connectingend arranging body 30 with respect to the measuring ends 44 _(j) of themeasuring device 40. For the movement speed thereof, it is moved at aspeed that is calculated based on the stable light receivable time, thefluorescence lifetime, and the number (twelve in this example) ofexclusive regions 20 _(i). Consequently, the receiving of light from alltwelve PCR tubes 231 _(i) within the stable light receivable timebecomes completed.

In step S16, the measurement control portion 61 determines the moment ofeach optical connection between the optical fibers (bundles) 33 _(i) ofthe linking portions 31 _(i) and the first measuring end and the secondmeasuring end of the measuring end 44, and instructs the receiving oflight to the measuring device 40 for example.

This measurement is executed with respect to cycles in which exponentialamplification is performed, and an amplification curve is obtained basedon the measurement, and various analyses are performed based on theamplification curve. At the time of the measurement, the measurementcontrol portion 61 heats the heater 37 built into the light guide stage32 and prevents the condensation on the sealing lid 251, and a clearmeasurement can be performed.

FIG. 11 is a perspective view of the front side of a nozzle head 500 ofa light measurement apparatus for a reaction vessel according to asecond exemplary embodiment of the present invention, and a perspectiveview showing a portion thereof cut away. FIG. 12 is a perspective viewshowing enlarged, the portion of FIG. 11 shown cut away.

As shown in FIG. 11, in this example, unlike the light measurementapparatus for a reaction vessel according to the first exemplaryembodiment, the PCR tubes 236 _(i), which represent the reaction vessel,have a vessel group in which three or more rows of twelve each arearranged.

There are no large differences with the first exemplary embodiment withrespect to the section of the nozzle head 500 related to a dispensingdevice, which includes the nozzles, and the section related to thetraversable nozzle, the transfer mechanisms of the nozzle head, and thearranging body transfer mechanism, and they are omitted from thedescriptions. The nozzle head 500 has: a light guide stage 320; twelvelinking portions 310 _(i) provided on the light guide stage 320; opticalfibers (bundle) 33 _(i) that extend from the linking portions 310 _(i)on the rear side; a connecting end arranging body 300; a measuringdevice 400 having a measuring end comprising six types of specificwavelength measurement devices that are aligningly mounted on the lightguide stage 320; and a sealing lid transporting body 125.

The light guide stage 320 according to the second exemplary embodimenthas a linking portion arranging body 322, in which two or more (twelvein this example) linking portions 310, that are simultaneously linkablewith two or more (twelve in this example) PCR tubes 236 _(i)representing reaction vessels, are arranged, that is movable in thehorizontal direction (the X axis direction in this example) with respectto the light guide stage 320. Furthermore, by means of the movement ofthe linking portion arranging body 322, without moving the light guidestage 320, it is linkable with more PCR tubes 236 _(i) representingreaction vessels (three rows of reaction vessels with twelve per row inthis example) than the number of reaction vessels (twelve in thisexample) that are simultaneously linkable by the linking portionarranging body 322.

The light guide stage 320 has a horizontal plate 320 a, a vertical plate320 b, and a triangular-shaped support side plate 320 c. The horizontalplate 320 a of the light guide stage 320, according to the arrangementof the linking portions 310, arranged on the linking portion arrangingbody 322, is etchingly provided with two or more, or twelve in thisexample, long holes 321 _(i) that correspond to shielding regions.

The measurement device 400 is mounted fixed to the upper edge of thevertical plate 320 b of the light guide stage 320. Therefore, since thelight guide stage 320 is stationary at the time of receiving light, themeasurement device 400 is immovably provided with respect to thereaction vessel and the light guide stage 320.

The optical fibers (bundle) 33 _(i) having the end of the linkingportion 310 _(i), separate midway into optical fibers for receivinglight (bundle) 331 _(i) and optical fibers for irradiation (bundle) 332_(i). The optical fibers for receiving light (bundle) 331 _(i) connectto a second connecting end 341 _(i), and the optical fibers forirradiation (bundle) 332 _(i) connect to a first connecting end 342_(i), and are arranged as two paths along the Y axis direction on adownwardly facing horizontal plane, which represents an arrangingsurface on the lower side of the connecting end arranging body 300. Atthat time, the spacing between adjacent connecting ends on theserespective paths is such that they are integrated at approximately halfor one-third of the spacing of the linking portions for example. Thefirst connecting ends 342 _(i) are successively connectable with thefirst measuring ends of the measurement device 400, and the secondconnecting ends 341 _(i) are successively connectable with the secondmeasuring ends.

As shown in FIG. 12 or FIG. 13, the horizontal plate 320 a of the lightguide stage 320 is laminatingly provided with a thermal insulation plate323 formed by a resin and the like, a heater 370 provided on the lowerside of the thermal insulation plate 323 for preventing condensation ofthe sealing lids 253 _(i) by heating the sealing lids 253 _(i), and athermally conductive metallic plate 325 provided on the lower side ofthe heater 370. Reference symbol 238 is a housing hole that houses thePCR tubes 236 _(i) representing reaction vessels, and is piercinglyprovided in the cartridge vessel. Reference symbol 239 represents aliquid surface that is controlled at a fixed height within the PCR tubes236 _(i) representing reaction vessels. Reference symbol 291 is atemperature controller for PCR.

The long holes 321 _(i) that are etchingly provided in the horizontalplate 320 a reach the metallic plate 325. Holes 326 for lighttransmission that are the same size as the apertures are piercinglyprovided above the apertures of the PCR tubes 236 _(i) representingreaction vessels, of the metallic plate 325 on the bottom of the longholes 321 _(i), and are optically communicated with the bottom of thelong holes 321 _(i).

The linking portions 310 _(i) provided on the linking portion arrangingbody and the ends of the optical fibers (bundle) 33 _(i) provided in theinterior are, as a result of approaching the sealing lids 253 _(i),linked with the PCR tubes 236 _(i) representing reaction vessels.

FIG. 14 is a drawing showing the various sealing lids 254 _(i) to 257_(i) according to the second exemplary embodiment, that are mountable onthe reaction vessel.

In FIG. 14A, the sealing lid 253 _(i) has: a cover plate 253 _(i) a thatcovers the aperture 236 _(i) a of the PCR tube 236 _(i) representing areaction vessel; a central portion 253 _(i) c that is formed at thecenter of the cover plate 253 _(i) a and thinner than the periphery, andhas an increased light transmittance; and a clamp 253 _(i) b comprisinga double annular wall that is provided such that it encloses the centralportion 253 _(i) c and protrudes on the lower side, that represents amounting portion that is mountable to the outer edge portion 236 _(i) bof the aperture of the reaction vessel.

The sealing lid 254 _(i) shown in FIG. 14B is formed thick in a convexlens form having a curved surface that expands from a central portion254 _(i) c toward the vessel exterior. Consequently, the light that isgenerated within the reaction vessel is made to converge at the end ofan optical fiber, or the excitation light from the optical fiber is madeto converge at the liquid surface and the like, and the light can beefficiently collected.

The sealing lid 255 _(i) shown in FIG. 14C is formed in a convex lensform having a curved surface that expands from a central portion 255_(i) c toward the vessel exterior, and consequently, the effectsdemonstrated in FIG. 14B are achieved.

The sealing lid 256 _(i) shown in FIG. 14D is formed thick such that ithas a curved surface that expands from a central portion 256 _(i) ctoward the vessel exterior. Consequently, the effects demonstrated inFIG. 14B are achieved.

The sealing lid 257 _(i) shown in FIG. 14E is formed such that it has acurved surface that expands from a central portion 257 _(i) c toward thevessel exterior, and consequently, the effects demonstrated in FIG. 14Bare achieved.

FIG. 15 shows a sealing lid transporting body 125 according to thesecond exemplary embodiment.

The sealing lid transporting body 125 is one having: a prismaticsubstrate 128 that is movable in the X axis direction with respect tothe vessel group 20, which has at least three rows of PCR tubes 236 _(i)representing reaction vessels of twelve per row; one or two or more(twelve in this example) grippers 127 _(i) arranged on the prismaticsubstrate 128 according to the arrangement of the reaction vessels thatgrip the cover plate such that, with respect to the sealing lid 253 _(i)(to 256 _(i)), the lower side is exposed in a state in which themounting portion is mountable to the reaction vessel; and a bottom plate126 that is mounted on the lower side of the prismatic substrate 128.

As shown in the cross-sectional view of FIG. 16 and the perspective viewas viewed from the lower side of FIG. 17, the grippers 127 _(i) have acavity 124 _(i) that is cut out from the prismatic substrate 128 in anapproximate semicircular column shape such that most of the cover plate253 _(i) a of the sealing lid 253 _(i) is housable. Furthermore, thebottom plate 126 is provided with a semicircular hole shaped notchportion 129 _(i) such that the clamp 253 _(i) b, which represents themounting portion of the sealing lid 253 _(i), is exposable on the lowerside.

Next, the processing operation using the nozzle head 500 according tothe second exemplary embodiment is described.

Among the processes described in the first exemplary embodiment, theseparation and extraction process is omitted, and step S′12 to stepS′16, which correspond to nucleic amplification and measurementprocesses, are described.

In step S′12, a new dispensing tip 211 _(i) is mounted on the nozzle 71_(i), the solution containing nucleic acids and the like, which ishoused within the eighth liquid housing part is aspirated, transportedto the PCR tube 236 _(i) representing a reaction vessel in which thesolution for amplification 234 _(i) is housed beforehand, and dischargedand introduced into the vessel. As a result of moving the nozzle head500 by means of the nozzle head transfer mechanism 51, the sealing lids253 _(i) from the sealing lid housing part in the sealing lidtransporting body 125 in which twelve sealing lids 253 _(i) are housed,are simultaneously housed in the cavity 124 _(i) of the grippers 127_(i), and gripped.

Since the sealing lid transporting body 125 gripping the sealing lid 253_(i) is linked with the light guide stage 320, then by using the stage Zaxis transfer mechanism 35 and moving it somewhat upwardly together withthe stage 320 and then moving it in the X axis direction, and bytransporting it to above the PCR tubes 236 _(i)representing reactionvessels and lowering it, the twelve sealing lids 253, are sealed bymounting the clamps 253 _(i) b, which are exposed on the lower side fromthe sealing lid transporting body 125, to the PCR tubes 236 _(i). In thesame manner, the rows of the twenty four reaction vessels of the othertwo rows are successively sealed by the sealing lids.

In step S′13, due to an instruction by the measurement control portion61, as a result of instructing the nozzle head transfer mechanism 51 andmoving the nozzle head 500 along the X axis, the light guide stage 320is moved such that it covers the thirty six reaction vessels of thethree rows, on which the sealing lids are mounted.

In step S′14, due to an instruction by the nucleic acid processingcontrol portion 63, the temperature controller 29 instructs atemperature control cycle by real-time PCR, such as a cycle in which thePCR tubes 231 _(i) are heated for five seconds at 96° C. and heated forfifteen seconds at 60° C., to be repeated forty nine times for example.

In step S′15, when temperature control at each cycle is started by thenucleic acid processing control portion 63, the measurement controlportion 61 determines the start of the elongation reaction process ateach cycle, moves the linking portion arranging body 322 provided on thelight guide stage 320 over the light guide stage 320, indirectly linksthe respective linking portions 310 _(i) that are inserted into the longholes 321 _(i) provided on the light guide stage 320 via the reactionvessels and the sealing lids 253 _(i), and successively receives thelight from the reaction vessels while irradiating excitation light fromthe measurement device to the interior of the reaction vessels. At thesame time, the continuous or intermittent movement of the connecting endarranging body 300 with respect to the respective measuring ends 44 _(j)of the measurement device 400 is instructed. The movement speed thereofis such that movement is performed at a speed that is calculated basedon the stable light receivable time, the fluorescence lifetime, thenumber (three rows of twelve reaction vessels per row in this example)of reaction vessels of the exclusive regions 20 _(i) that are measurableby the light guide stage 320, and the like. Consequently, by moving thelinking portion arranging body 322 over the light guide stage 320 withinthe stable light receivable time, in this example, measurements can beperformed in parallel with respect to thirty six reaction vessels ofthree rows, with twelve per row.

In step S′16, the measurement control portion 61 determines the momentof the respective optical connections between the optical fibers(bundle) of the linking portions 310 _(i) and the first measuring endand the second measuring end of the measuring end 44, and instructs theirradiation of excitation light and the receiving of light to themeasurement device 400.

FIG. 18 is a drawing showing an example of the position of the opticalfiber front ends for receiving light and for irradiation provided to thelinking portion in a case where the linking portion is linked at alocation other than the aperture of the PCR tube 236 _(i) representing areaction vessel. FIG. 18A is a drawing showing a case where the opticalfibers (bundle) for receiving light 331 _(i) are in the vicinity of theouter bottom portion of the reaction vessel, and the optical fibers(bundle) for irradiation 332 _(i) are in the vicinity of the outer wallof the reaction vessel. FIG. 18B is a drawing showing a case where theoptical fibers (bundle) for receiving light 331 _(i) and the opticalfibers (bundle) for irradiation 332 _(i) are in the vicinity of theouter wall of the reaction vessel. FIG. 18C is a drawing showing a casewhere the optical fibers (bundle) for receiving light 331 _(i) and theoptical fibers (bundle) for irradiation 332 _(i) are in the vicinity ofthe outer bottom portion of the reaction vessel. These are onlyexamples, and cases where they are joined with the reaction vessel bymaking contact, and the like, in place of being in the vicinity are alsopossible.

FIG. 19 represents a block-diagram of a light measurement apparatus fora reaction vessel 110 according to a second embodiment of the presentinvention. Since the same reference symbols as the reference symbolsused in the first embodiment represent the same objects or similar (onlydiffering by size) objects, the descriptions thereof are omitted.

The light measurement apparatus for a reaction vessel 110 according tothe second embodiment differs from the light measurement apparatus for areaction vessel 10 according to the first embodiment in the aspect thatthe nozzle head 150 thereof has a light guide stage 132 that isdifferent from the light guide stage 32. The light guide stage 132according to the second embodiment differs from the light guide stage 32according to the first embodiment in the aspects that it has a plurality(twelve in this example) of linking portions 131 _(i) in which the frontends of optical fibers, which represent two or more light guideportions, which have a flexibility, that optically connect with theinterior of the PCR tubes 231 _(i), and an optical element forcollecting light are provided in the interior, and the heat source ofthe heater 137, which represents a heating portion for heating thereaction vessels, is provided not to the light guide stage 132, but tothe vessel group 120 or the stage.

Further, it differs in the aspects that the sealing lids 251 _(i) aretransported not by the nozzles 71 _(i) but by fitting to the linkingportions 131 _(i), and are detached from the linking portions by meansof a dedicated sealing lid detaching mechanism 39. Therefore, thesealing lid control portion 167, and therefore, the nucleic acidprocessing control portion 163 and the CPU+program 160 differ from thedevice 10 according to the first embodiment.

The vessel group 120 is one in which twelve exclusive regions 120 _(i)(i=1, . . . , 12), wherein the longitudinal direction thereof is alongthe X axis direction and housing parts are arranged in a single rowform, are arranged in the Y axis direction for example. The respectiveexclusive regions 120, have: a reaction vessel group 23 _(i); a liquidhousing part group 27 _(i); a sealing lid housing part 25 _(i) thathouses sealing lids 251 _(i), which have transparency, that aredetachably mounted on the linking portions 131 _(i) provided to thelight guide stage 132; and housing parts for tips and the like 21 _(i).

The reaction vessel 23 _(i), the temperature controller 29, and theheater 137 correspond to the reaction vessel control system 90.

FIG. 20 is a cross-sectional view primarily showing, within the nozzlehead 150 according to a first exemplary embodiment of the secondembodiment, the transfer mechanism and the suction-discharge mechanism.

Here, since the diameter of the linking portion 131 _(i) is thicker thanthe nozzle 71 _(i), the sealing lid 251 _(i) to be mounted on the PCRtube is transported by the linking portion 131 _(i). Consequently, byutilizing the transfer mechanism of the magnet 571 of the magnetic forcepart 57, a sealing lid detaching mechanism 39 is provided that has acomb-shaped detaching member 391 in which a notch portion, which has asemicircular-shaped arch that is approximately equivalent to thediameter of the twelve linking cylinders provided such that they canapproach and separate with respect to the linking portion 131 _(i), isarranged. In the present exemplary embodiment, since the detachment ofthe sealing lids can be performed by utilizing existing mechanisms, theexpansion of the apparatus scale, and increases in complexity, can beprevented.

FIG. 21 is a drawing showing a reaction vessel control system 901according to the first exemplary embodiment of the second embodiment anda state in which, to the apertures of the reaction vessel group, towhich the PCR tubes 231 _(i) representing a plurality (twelve in thisexample) of reaction vessels of the reaction vessel control system 901are provided, the linking portions 131, (here, i=1 for example)protruding on the lower side from the horizontal plate 132 a of thelight guide stage 132 are indirectly linked with the PCR tubes 231 i viathe sealing lids 251 i, which have transparency, that are mounted on theapertures of the PCR tubes 231 _(i) in the exclusive regions 120 _(i).As a result of the linking portions 131 _(i) fitting within theindentation for linking 253 _(i) of the sealing lids 251 _(i), they arelinked with the PCR tubes 231 _(i).

As shown in FIG. 21, the linking portion 131 _(i) is indirectly linkedwith the PCR tube 231 _(i) via the sealing lid 253, and has anapproximately cylinder-shaped linking cylinder 131 a _(i) that isprotrudingly provided further in the downward direction than thehorizontal plate 132 a of the light guide stage 132. Furthermore, acircular hole 131 b _(i) having an aperture of a size corresponding tothe liquid surface of the liquid that is housed in the narrow-mouthedpiping part is piercingly provided in the center portion of the bottomplate of the linking cylinder 131 a _(i), and the periphery of thebottom plate is provided with a circular edge portion 131 d _(i) that isprotrudingly provided below it. Consequently, the adhesion of thelinking portion and the sealing lid is prevented. A spherical ball lens381 _(i) that has a diameter corresponding to the inner diameter of thelinking cylinder is loosely inserted within the linking cylinder 131 a_(i) and mounted on the circular hole 131 b _(i). At a predetermineddistance above the ball lens 381 _(i), an optical fiber 33 _(i), inwhich the end is positioned and is covered by a resin-made ferrule 131 c_(i) that passes through the horizontal plate 132 a and reaches theexterior, is provided. The linking cylinder 131 a _(i), the circularhole 131 b _(i), the ball lens 381 _(i), and the optical fiber 33 _(i)bundle are arranged on the same axis in the interior of the linkingcylinder 131 a _(i).

As shown in FIG. 21, the reaction vessel control system 901 has: PCRtubes 231 _(i) that represent reaction vessels, in which targetsolutions of DNA having a target base sequence, and the like, are storedand reactions, such as amplification, are performed; a heater 137; and atemperature controller 291 _(i) for PCR. The heater 137 is laminatinglyprovided with a heating block 137 c comprising an aluminum plate havinga high thermal conductivity, a sheet heater 137 a, and a heat insulator137 b. Twelve through holes 137 d _(i) that house and retain a plurality(twelve in this example) of PCR tubes 231 _(i) are piercingly providedin the same heater 137, and the wide-mouthed piping parts 235 _(i) aresupported by the heating block 137 c.

The temperature controller 291 for PCR has: a block for temperaturecontrol 292 _(i) that makes contact with, and is housable in, thenarrow-mouthed piping part 233 _(i) of the PCR tube 231 _(i), whichrepresents the reaction vessel; a Peltier element 293 _(i); and a heatsink 294 _(i).

The narrow-mouthed piping part 233 _(i) of the PCR tube 231 _(i) has alower side wall section 233 a _(i) of the section in which the block forPCR 292 _(i) is making contact and is provided. Furthermore, it has anupper side wall section 235 a _(i) provided on the upper side leaving aspacing with the lower side wall section 233 a _(i) that corresponds tothe wall section of the wide-mouthed piping part 235 _(i) that makescontact with the block for heating 137 c of the heater.

According to the present exemplary embodiment, firstly, by means of aninstruction by the sealing lid control portion 167 (the CPU+program160), the nozzle head transfer mechanism 51 is instructed and followingmovement of the respective linking portions 131 _(i) of the light guidestage 132 to the sealing lid housing parts 25 _(i), the stage Z axistransfer mechanism 35 is instructed and the sealing lids 251 _(i) arefitted and mounted to the linking portions 131 _(i). Next, by fittingthe apertures of the predetermined PCR tubes 231 _(i) with the sealinglids 251 _(i), the linking portions 131 _(i) are simultaneously linkedwith the PCR tubes 231 _(i).

Next, according to the temperature control by the temperature controller29 as a result of an instruction by the measurement control portion 161,in the case of PCR, by controlling the heater 137 such that the upperside wall section 235 a _(i) is heated at a fixed temperature (100° C.for example) that is several degrees, or preferably approximately 5° C.,higher than the maximum predetermined temperature (94° C. for example),the sealing lid 251 _(i) fitted to the wide-mouthed piping part 235 _(i)of the PCR tube 231 _(i) is heated, and condensation of the sealing lidcan be prevented. At that time, the upper side wall section 235 a _(i)is separated by a fixed spacing from the lower side wall section 233 a_(i), in which temperature control is performed, and the upper side wallsection 235 a _(i), which has a smaller surface area than the lower sidewall section, is heated by bringing the heat source into contact or intoits vicinity. Consequently, the effect of heating the upper side wallssection 235 a _(i) is such that the lower surface of the sealing lid 251_(i), which is provided at a position near the upper side walls section235 a _(i), is heated, and condensation can be prevented.

On the other hand, since the linking portion 131 _(i) is only makingcontact with the upper side of the sealing lid 251 _(i) via the circularedge portion 131 d _(i), the effect of heating is not as much as withrespect to the sealing lid 251 _(i). In the same manner, the lower sidewall section 233 a _(i) is temperature controlled to the predeterminedtemperature using a Peltier element having a heating and coolingfunction, and furthermore, measurements are simultaneously performed.After completion of the measurement, then by means of an instruction bythe sealing lid control portion 167, the linking portion 131 _(i) ismade to approach using the detaching member 391, and then by upwardlymoving the light guide stage 132 by means of the stage Z axis transfermechanism 35, the sealing lid 251 _(i) is detached from the linkingportion and while remaining on the PCR tube 231 _(i), the linkingportion is moved and the linking is released.

FIG. 22 is a drawing showing a second exemplary embodiment, andrepresents a linking portion 131 _(i) in which, in place of the balllens 381 _(i), a drum lens 382 _(i) having a lens diameter correspondingto the inner diameter of the linking cylinder 131 a _(i) is looselyinserted within the linking cylinder 131 a _(i) and mounted on thecircular hole 131 b _(i), and is provided such that light is collectedat the end of the optical fiber 33 _(i).

FIG. 23 is a drawing showing a third exemplary embodiment, andrepresents a linking portion 131 _(i) in which, in place of the balllens 381 _(i) and the like, an aspheric surface lens 383 _(i) having alens diameter corresponding to the inner diameter of the linkingcylinder 131 a _(i) is loosely inserted within the linking cylinder 131a _(i) and mounted on the circular hole 131 b _(i), and is provided suchthat light is collected at the end of the optical fiber 33 _(i).Reference symbol 391 represents a comb-shaped detaching member of thesealing lid detaching mechanism 39, and shown is a state in which it isin the vicinity of, or making contact with, the linking portion 131_(i). In this state, by raising the linking portion 131 _(i), thesealing lid 251 _(i) engages with the sealing lid detaching member 391and is detached from the linking portion 131 _(i), but remains stillmounted on the PCR tube 231 _(i). Furthermore, the respective lenses 381_(i) to 383 _(i) may be made to be loosely mounted within the linkingcylinder 131 a _(i) by installing a tube-shaped frame from the upperside.

FIG. 24 is a drawing in which PCR tubes 231 _(i) (FIG. 24A) are housedand retained in twelve through holes 137 d provided in a heater 137(FIG. 24B) according to the present embodiment, with the heater 137 thenheated to a predetermined temperature. The top surface temperature ofthe sealing lid is measured at the time the heating temperature of theheater thereof is increased, and the temperature at which condensationoccurs, and the temperature at which condensation is eliminated, aremeasured.

As shown in FIG. 24A, the apertures of the PCR tubes 231 _(i) are sealedby a sealing lid 259, which has a transparency. The diameter of theupper side of the sealing lid is 14 mm, the height of the mountingportion of the sealing lid 259 is 8.6 mm, and the height from the baseof the PCR tube to the upper side of the sealing lid 259 is 21.4 mm.FIG. 24C is a drawing showing the actual measured temperature at the topsurface of the sealing lid with respect to the set temperature of theheater.

In step S101, 25 μL of distilled water is dispensed into thenarrow-mouthed piping parts of the respective PCR tubes 231 _(i), and asshown in FIG. 24A, the apertures of the wide-mouthed piping parts aresealed by the sealing lid 259, and the PCR tubes 231 _(i) are retainedin the twelve through holes of the heater 137. The top surfacetemperature of the sealing lid 259 is measured by opening a small hole(diameter: 0.5 mm) in the sealing lid 259, inserting a thermocouple 900,and installing it on the inside of the top surface of the sealing lid259.

In step S102, the temperature of blocks for PCR 292 _(i), which areprovided such that the narrow-mouthed piping parts of the PCR tubes 231_(i) are making contact, are set to 95° C., and the presence ofcondensation on the top surface of the sealing lid is visually observed.At this time, the set temperature of the heater 137 is gradually raisedfrom 100° C. to 115° C., and the set temperature at which condensationis eliminated is confirmed. Reference symbol 902 represents the positionof a thermistor, and a thermocouple is installed in the vicinitythereof.

In a separate experiment, the temperatures were higher, in blockdivision units of the heater 137, in the order of lanes 1 to 4, lanes 9to 12, and lanes 5 to 8. When the top surface temperature of the sealinglid was measured for lane 1, which had the lowest temperature, and forlane 6, which had the highest temperature, the top surface temperatureof the sealing lid of lane 1 was 13° C. lower than the set temperature,and the top surface temperature of the sealing lid of lane 6 wasvirtually the same as the set temperature. When the set temperature was113° C., condensation was resolved at all lanes, and it can be inferredthat the top surface temperature at lane 1 at this time wasapproximately 100° C.

FIGS. 25A, 25B and 25C are drawings showing, when the material of thelinking portions 131 _(i) is stainless steel, or when it is a resin witha lower thermal conductivity than stainless steel (PEEK), an improvementto the anti-fogging effect when they are linked with the PCR tubes 231_(i). FIG. 25A is a drawing showing a PCR tube 231 to which a linkingportion 131 is linked. FIGS. 25B and 25C respectively show thetemperature-fluorescence intensity characteristics when a stainlesssteel-made linking portion and a PEEK-made linking portion are used. Thevertical axis represents the fluorescence intensity.

In this experiment, in step S201, 20 μL of a fluorescent reagent (0.25μL ROX) is dispensed into the PCR tubes 231 _(i) representing thereaction vessels, the apertures of which are sealed by the sealing lid259, and the equivalent of twelve lanes are set to the temperaturecontrol block. Then, the linking portion 131 is mounted to theindentation section of the sealing lid. Here, the linking portions 131_(i) have an outer diameter of 10 mm.

In step S202, the block temperature is raised from 60° C. to 95° C. bymeans of the temperature control block, and the fluorescence intensityis measured for the respective lanes every 1° C. Consequently, incontrast to a marked decrease in the fluorescence intensity beingobserved at a PCR block temperature of approximately 80° C. or higherwhen a stainless steel-made linking portion is used, this is improvedwhen a PEEK-made linking portion is used. Here, in comparison to thethermal conductivity (approximately 16.7 W/(m·K)) of stainless steel(SUS303), the thermal conductivity of PEEK of approximately 0.25 W/(m·K)is very low. Therefore, it is considered that the anti-foggingproperties are improved as a result of a decrease in the heat releasefrom the heater 137 that occurs by transmission to the linking portion,and the heating effect toward the sealing lid thereby improving.

FIG. 26 is a drawing showing the thermal effects of heat transfer fromthe heater 137 imparted on an optical fiber 33 representing the lightguide portion, which is provided within the linking portion 131 andsurrounded by a ferrule 131 c.

In conclusion, the temperature within the ferrule is a maximum of 41° C.at the time of execution of temperature control by a thermal cycle. Thisis sufficiently lower than the heat resistance temperature of 70° C. ofthe optical fiber 33 representing the light guide portion, and it isconsidered that heat transfer from the heater imparts no effects on thedegradation of the optical fiber.

The experiment was performed using a thermo-recorder (KEYENCENR-250(K05003)), an extra fine thermocouple 01-K manufactured byNinomiya Electric Wire Co., Ltd. as a temperature sensor, and a ferrule(front end outer diameter: 3.5 mm, length: 32.5 mm) not containing afiber.

The thermocouple was inserted into the ferrule. A Teflon (registeredtrademark) tube was pressed into the ferrule, and one of thethermocouples was pressed and fixed against the inner wall of theferrule. Two such ferrules were prepared and installed within thelinking portion 131. The linking portions were mounted onto lane 6,which had the highest temperature on the heater 137, and lane 1, whichhad the lowest temperature. Then, after PCR temperature control to 95°C. (2 min), a thermal cycle that repeats 50 times a cycle from 60° C.(30 sec) to 95° C. (5 sec) was executed, and the temperature changewithin the ferrule was recorded. The temperature of the heater 137 wasset to 113° C., at which condensation was eliminated.

In FIG. 26, (1) represents room temperature, (2) represents thetemperature within the apparatus, (3) represents the temperature of lane1, and (4) represents the temperature of lane 6.

As a result of this experiment, the temperature within the ferrule 131 cat completion of the thermal cycle had risen by approximately 2° C.compared to the start. Therefore, the temperature within the ferrule atcompletion of the thermal cycle was a maximum of approximately 41° C.(steady state). Consequently, this temperature is sufficiently lowerthan the heat resistance temperature of 70° C. of the optical fibers 33_(i) of the light guide portions, and it is considered that heattransfer from the heater 137 imparts no effects on the degradation ofthe optical fibers 33 _(i) within the linking portions 131 _(i).

FIG. 27 is a drawing showing extracted two adjacent specific wavelengthmeasurement devices 140 ₁ and 140 ₂ among the six specific wavelengthmeasurement devices 140 _(j) (j=1, . . . , 6) that correspond to theplurality of types of measurement devices 140 according to a fourthembodiment of the present invention.

The specific wavelength measurement devices 140 ₁ and 140 ₂ are directlyor indirectly linkable with the two or more reaction vessels 231 _(i),and are provided corresponding to two or more linking portions 131 _(i),to which are provided the front ends of two or more light guide portions133 _(i), which have a flexibility, that optically connect with theinterior of the linked reaction vessels 231 _(i). The configuration issuch that the two or more connecting ends 134 _(i), which are providedwith the rear ends of the light guide portions that have the front endsprovided to the linking portions 131 _(i), are provided in the vicinityof, or making contact with, the arranging surface of the connecting endarranging body 130, which maintains them along the predetermined path ina specifiable array, and have one or two or more measuring ends 142 ₁and 143 ₁ (142 ₂ and 143 ₂) that are successively optically connectablewith the respective connecting ends 134 _(i) along the predeterminedpath. They are thus able to receive the light based on the optical statewithin the reaction vessels by means of an optical connection betweenthe connecting ends 134 _(j) and the measuring ends 142 _(i) and 143_(i).

There are provided corresponding to the respective specific wavelengthmeasurement devices 140 ₁ and 140 ₂, at the very least between adjacentspecific wavelength measurement devices, modulators 48 ₁ and 48 ₂ thatmodulate the intensity of the light of the specific wavelengths Λ1 andΛ2 to be received by the respective specific wavelength measurementdevices at the mutually different predetermined frequencies A and B, andtuners 49 ₁ and 49 ₂ that perform tuning from the received light to thefrequency of the light intensity of the modulated specific wavelengthsΛ1 and Λ2.

In a case where the light based on the optical state is a fluorescentlight, the respective specific wavelength measurement devices 140 ₁ and140 ₂ are provided with irradiation sources 146 ₁ and 146 ₂ thatirradiate the predetermined excitation light λ1 and λ2 that excitefluorescent light of the corresponding specific wavelengths or specificwavelength bands Λ1 and Λ2, and light receiving portions 147 ₁ and 147 ₂that are able to receive fluorescent light of the specific wavelengthsor the specific wavelength bands Λ1 and Λ2. Generally, the wavelength ofthe excitation light λ is shorter than the specific wavelength Λ.

The irradiation sources 146 ₁ and 146 ₂ are provided with light emittingelements 1464 ₁ and 1464 ₂, such as an LED or an incandescent bulb, thatemit light containing the corresponding excitation lights λ1 and λ2, andan excitation light λ1 optical filter 1468 ₁ and an excitation light λ2optical filter 1468 ₂ that extract and transmit the excitation lights λ1and λ2 from the emissions of the light emitting elements 1464 ₁ and 1464₂.

The light receiving portions 147 ₁ and 147 ₂ are provided with aspecific wavelength Λ1 optical filter 1477 ₁ and a specific wavelengthΛ2 optical filter 1477 ₂ that extract and transmit light of thecorresponding specific wavelengths or the specific wavelength bands Λ1and Λ2, and light receiving elements 1474 ₁ and 1474 ₂, such as aphotodiode, a phototransistor, a photomultiplier tube, a photoelectriccell, or a photoconductive cell, which convert the light transmitted bythe specific wavelength Λ1 optical filter 1477 ₁ and the specificwavelength Λ2 optical filter 1477 ₂ into a corresponding electricalsignal.

Consequently, the modulators 48 ₁ and 48 ₂ have a frequency Aoscillating voltage supply circuit 148 ₁ and a frequency B oscillatingvoltage supply circuit 148 ₂ representing the excitation lightmodulation portions, which supply to the irradiation sources 146 ₁ and146 ₂ voltages that oscillate with a predetermined waveform at mutuallydifferent predetermined frequencies A and B, which are voltages fordriving the light emitting elements of the irradiation sources 146 ₁ and146 ₂. The modulators 48 ₁ and 48 ₂ also include the irradiation sources146 ₁ and 146 ₂, and in addition, the reaction vessels 231 _(i) in whichthe fluorescent compound is housed.

On the other hand, the tuners 49 ₁ and 49 ₂ tune the electrical signalsconverted from the light received by the light receiving portions 147 ₁and 147 ₂ to the frequency A or the frequency B by passing them througha frequency A bandpass filter circuit or a frequency B bandpass filtercircuit. Then, of the fluorescent light emitted based on the excitationlights λ1 and λ2 from the specific wavelength measurement devices 140 ₁and 140 ₂, the light intensity data (electrical signals) for thefrequency A or the frequency B that is received is extracted, whichprevents crosstalk between adjacent specific wavelength measurementdevices 140 ₁ and 140 ₂, and measurements with a high reliability can beperformed.

FIG. 28, which corresponds to FIG. 27, shows the movement directionbetween six pairs of measuring ends 142 _(j) and 143 _(j) of theplurality of types (six types in this example) of specific wavelengthmeasurement devices 140 _(j) (j=1, . . . 6), and the connecting ends 134_(i) of the connecting end arranging body 130.

In FIGS. 29A, 29B, 29C, 29D, and 29E the respective modulatorscorresponding to the six pairs of specific wavelength measurementdevices 140 _(j) (j=1, . . . 6) shown in FIG. 28 are set to therespective frequencies A to F (A, B, C, D, E, F)=(1 kHz, 4 kHz, 7 kHz, 7kHz, 4 kHz, 1 kHz). In a case where the intensity of the excitationlight is modulated as a blinking signal, a non-reflective sheet housedwithin the reaction vessels is irradiated instead of a sample, and datathat is tuned to the respective frequencies from the electrical signalof the light received at a sampling rate of 5 ms is extracted. Then, thecrosstalk between the specific wavelength measurement devices 140 _(j)was measured by measuring the intensity (data that corresponds tointensity data of the received fluorescent light) of the correspondingexcitation light. Here, the intensity of the excitation light isoverwhelmingly higher (on the order of several hundred times) than theintensity of the fluorescent light. Furthermore, the wavelength of theexcitation light is short compared to the wavelength of the fluorescentlight that is excited. However, if several types of fluorescent lightare used, there are cases where the wavelength or the wavelength band ofthe excitation light overlaps with the wavelength or the wavelength bandof the fluorescent light used in an adjacent measurement device thatdoes not support the excitation light thereof. In such a case, there isa concern of the excitation light not being able to be excluded at thetime the fluorescent light is received, generating crosstalk between themeasurement devices.

In the case of FIG. 29A (1), only the specific wavelength measurementdevices 140 ₁, 140 ₂, and 140 ₃ provided on the substrate 1 are set withmutually different frequencies A, B, and C (=1 kHz, 4 kHz, 7 kHz), andfor the case where the respective excitation lights λ1, λ2, and λ3 areirradiated, there is shown the intensities of the excitation lightsreceived by the respective measurement devices as a result of leaks,scattering, reflection, and the like. In this case, as shown in thegraph of (1), light of a constant intensity is respectively obtained.Therefore, crosstalk from the respective adjacent specific wavelengthmeasurement devices was not observed.

In the case of FIG. 29B (2), all of the specific wavelength measurementdevices 140 ₁, 140 ₂, and 140 ₃ provided on the substrate 1 and 140 ₄,140 ₅, and 140 ₆ provided on the substrate 2 are set with thefrequencies A to F as described above. Then, the respective excitationlights λ1, λ2, and λ3 are irradiated, and shown are the intensities ofthe excitation lights measured at the specific wavelength measurementdevices 140 ₁, 140 ₂, and 140 ₃ provided on the substrate 1. In thiscase, as shown in the graph of (2), the specific wavelength measurementdevice 140 ₃ is such that crosstalk of the excitation light exists withthe adjacent specific wavelength measurement device 140 ₄ due to theexcitation light being modulated at the same frequency of 7 kHz, and itis observed that there is a fluctuation in the intensity resulting froma combination of an offset in the phases of the modulated waveforms(blinking type) of the two devices.

In the case of FIG. 29C (3), shown is a case where all of the specificwavelength measurement devices 140 ₁, 140 ₂, and 140 ₃ provided on thesubstrate 1, and only the specific wavelength measurement device 140 ₆provided on the substrate 2 are driven, and a measurement is performedat the specific wavelength measurement devices provided on the substrate1. As shown in the graph of (3), crosstalk of the excitation light isnot observed since the spacing between measurement devices having thesame frequency is long.

In the case of FIG. 29D (4), shown is a case where all of the specificwavelength measurement devices 140 ₁, 140 ₂, and 140 ₃ provided on thesubstrate 1, and only the specific wavelength measurement device 140 ₅provided on the substrate 2 are driven, and a measurement is performedat the specific wavelength measurement devices provided on the substrate1. As shown in the graph of (4), crosstalk of the excitation light isnot observed.

In the case of FIG. 29E (5), shown is a case where all of the specificwavelength measurement devices 140 ₁, 140 ₂, and 140 ₃ provided on thesubstrate 1, and only the specific wavelength measurement device 140 ₄provided on the substrate 2 are driven, and a measurement is performedat the specific wavelength measurement devices provided on the substrate1. As shown in the graph of (5), the specific wavelength measurementdevice 140 ₃ is such that crosstalk of the excitation light exists withthe adjacent specific wavelength measurement device 140 ₄ due to theexcitation light being modulated at the same frequency of 7 kHz, and itis observed that there is a fluctuation in the intensity resulting froma combination of an offset in the phases of the modulated waveforms ofthe two devices.

In FIG. 30, shown is a case where, in accordance with 96-wellmicroplates that are utilized as an international standard, the linkingportions 131 are arranged with a 9 mm pitch corresponding to the pitchof 9 mm when the PCR tubes 231, representing the reaction vessels arearranged, and the spacing of the connecting ends 134 _(i) on thearranging surface of the connecting end arranging body 130 is integratedto 3.75 mm when the diameter of the optical fiber 133, which representsthe light guide portion, that is arranged on the connecting endarranging body 130 with one end thereof provided to the linking portion131, and the other end to the connecting end 134, is 1.5 mm. Even withsuch a spacing, as shown in FIG. 31, the data of the light received bythe measuring end 143 ₁ of a given specific wavelength measurementdevice 140 ₁ of the measurement device 140 identifies with certainty thelight from the eight connecting ends.

FIG. 32 is a drawing showing a connecting end arranging body 1300according to a fourth embodiment of the present invention, and themovement thereof The respective linking portions 131 ₂ to 131 ₁₂ areprovided with the front ends of light guide portion bundles 133 ₂ to 133₁₂ comprising a plurality of light guide portions. Among the light guideportion bundles, a portion of the rear ends of the light guide portionbundles is provided to a first connecting end of the connecting endarranging body 1300, and the remaining light guide portion bundles areprovided to a second connecting end of the connecting end arranging body1300. The predetermined path comprises a linear first path and a linearsecond linear path for example. The connecting end arranging body issuch that, with regard to the eleven pairs of first connecting ends andsecond connecting ends that are optically connected with the elevenlinking portions 131 ₂ to 131 ₁₂, and a single pair of third connectingends and fourth connecting ends that are optically mutually connected bymeans of a light guide portion 133 ₀, the eleven first connecting endsand the single third connecting end are arranged along the linear firstpath, and the eleven second connecting ends and the single fourthconnecting end are arranged along the linear second path, which isparallel to the first path leaving a predetermined spacing. In thisexample, for simplicity, the connecting end arranging body 1300 of thepresent embodiment has been achieved by removing the light guide bundle133 ₁ provided to the first pair of the first connecting end and thesecond connecting end of the connecting end arranging body 130 accordingto the embodiment described above, and further replacing it with thelight guide portion 133 ₀ that optically mutually connects between thethird connecting end and the fourth connecting end.

As a result of movement of the connecting end arranging body 1300, thefirst measuring ends 140 ₁ respectively provided to the measurementdevices 140 ₁ to 140 ₆ relatively move along the first path comprisingthe eleven first connecting ends and the single third connecting end,and the second measuring ends respectively provided to the measurementdevices 140 ₁ to 140 ₆ relatively move along the second path comprisingthe eleven second connecting ends and the single fourth connecting ends,respectively. The first measuring ends are optically connected with theirradiation source of the measurement device, and the second measuringends are connected with the light receiving portion of the measurementdevice. In contrast to a device in which the pairs of first measuringends and second measuring ends respectively provided to the measurementdevices 140 ₁ to 140 ₆ are integrally provided, as a result of movementof the connecting end arranging body 1300, for each pair, the firstmeasuring ends of the respective measurement devices successivelyconnect with the eleven first connecting ends and the single thirdconnecting end, and at the same time, the second measuring ends of therespective measurement devices successively connect with thecorresponding eleven second connecting ends and the single fourthconnecting end. Therefore, together with measurements of thefluorescence intensity by the eleven pairs of first connecting ends andsecond connecting ends, it becomes possible, by means of the single pairof the third connecting end and the fourth connecting end, for the lightreceiving portion to directly receive the excitation light via the lightguide portion 133 ₀ without it being diverted to the reaction vessels,and a genuine excitation light intensity can be measured.

FIG. 33A is a graph plotting, in a case where the temperature of thesurroundings of the LED has been intentionally raised (by stopping thecooling fan for example), the intensity of excitation light measured bythe light receiving portion using the pair of the third connecting endand the fourth connecting end, and data measured for the intensity offluorescent light obtained from the respective reaction vessels byirradiating excitation light using the eleven pairs of first connectingends and second connecting ends with respect to the reaction vessels viathe linking portions 131 ₂ to 131 ₁₂. In the case of this example, thecenter wavelength of the LED is 590 nm, and a modulated frequency of 7kHz is added to the excitation light of the LED circuit. Furthermore,distilled water is housed in the respective tubes, and after heating for5 min at 95° C., a temperature control of 95° C. for 5 sec and 60° C.for 30 sec is repeated for 80 cycles. FIG. 33B is a graph plotting thefluorescence intensities, which have been normalized by removing afluctuating component of the excitation light by dividing thefluorescence intensity by the measured intensity of the excitationlight.

As shown in FIG. 33A, together with the temperature increase of the LED,the intensity of the excitation light measured by the receiving portionusing the single pair of the third connecting end and the fourthconnecting end is gradually becoming weaker. Consequently, in that case,the nature of the fluctuations in the intensity of the excitation lightand the respective fluorescent light from 5 min to 80 min afterstabilization of the intensity is expressed as approximately 1.89% to1.37% as a CV value, in which the standard deviation is divided by theaverage value. In contrast, as shown in FIG. 33B, looking at the valueof the intensity of the fluorescent light divided by the intensity ofthe excitation light measured using the single pair of the thirdconnecting end and the fourth connecting end, the nature of thefluctuations in the intensity of the fluorescent light showsapproximately 0.40% to 0.63% as a CV value. Therefore, the fluctuationsin the fluorescence intensity are small. That is to say, it shows thatfluorescence intensities that have excitation light fluctuationsremoved, which are mainly determined by the fluorescent materialconcentration, are obtained.

FIG. 34 shows specific wavelength measurement devices 1400 ₁ and 1400 ₂in a case where, as tuners 49 ₁ and 49 ₂, lock-in amplifiers 1490 ₁ and1490 ₂ are used instead of bandpass filter circuits. In this case, thereference signal can utilize the outputs of the frequency A and Boscillating voltage supply circuits 148 ₁ and 148 ₂ of the modulators 48₁ and 48 ₂. Therefore, the configuration is simplified as a whole, andmeasurements with a high reliability can be performed.

The foregoing exemplary embodiments have been specifically described inorder to better understand the present invention, and they are in no waylimiting of other embodiments. Therefore, modifications are possiblewithin a scope that does not depart from the gist of the invention. Theconfigurations, shapes, materials, arrangements, numbers, and amounts ofthe nozzles, the dispensing tips, the punching tips, the vessel group,the exclusive regions thereof, the housing parts, the measuring ends,the measurement devices, the specific wavelength measurement devices,the suction-discharge mechanism, the transfer mechanism portion, themagnetic force part, the heating portion, the reaction vessels, thesealing lids, the light guide stage, the linking portions, the lightguide portions, the connecting ends, the connecting end arranging body,the linking portion arranging body, the nozzle head, the temperaturecontroller, the nozzle detaching mechanism, the sealing lid detachingmechanism, the modulators, the tuners, and the like, and the utilizedreagents and samples are also in no way limited by the examplesillustrated in the exemplary embodiments. Furthermore, although thenozzles were made to move with respect to the vessel group, it ispossible to also move the vessel group with respect to the nozzles.

Furthermore, in the foregoing descriptions, although the amplificationsolution was sealed using a sealing lid for the sealing of the reactionvessel for PCR, it may be made such that, in its place or incombination, it is sealed using a sealing liquid, such as mineral oil.Furthermore, in place of punching by mounting a tip for punching on thenozzles, it is possible to use a punching pin that is driven by thesuction-discharge mechanism. Moreover, although a real-time PCRmeasurement was described in the foregoing descriptions, it is in no waylimited to this measurement, and it can be applied to other variousmeasurements in which temperature control is performed. In the foregoingdescriptions, although a case where the measurement device is providedto a dispensing device was described, it is not necessarily limited tothis. Although only an optical system using optical fibers wasdescribed, it is possible to also employ an optical system using a lenssystem in the interior of the measurement device.

Furthermore, the apparatuses described in the respective exemplaryembodiments of the present invention, the components that form theseapparatuses, or the components that form these components, can beappropriately selected, and can be mutually combined by applyingappropriate modifications. The spatial representations within thepresent application, such as “above”, “below”, “interior”, “exterior”,“X axis”, “Y axis”, and “Z axis” are for illustration only, and are inno way limiting of the specific spatial directions or arrangements ofthe construction.

INDUSTRIAL APPLICABILITY

The present invention is related to fields in which the processing,testing, and analysis of nucleic acids, which primarily includes DNA,RNA, mRNA, rRNA, and tRNA for example, is required, and is related toindustrial fields, agricultural fields such as food, agriculturalproducts, and fishery processing, chemical fields, pharmaceuticalfields, health care fields such as hygiene, insurance, diseases, andgenetics, and scientific fields such as biochemistry or biology forexample. The present invention is, in particular, able to be used inprocessing and analysis that handles various nucleic acids, and thelike, such as PCR and real-time PCR.

BRIEF DESCRIPTION OF THE REFERENCE SYMBOLS

-   10, 110 Light measurement apparatus for reaction vessel-   20, 120 Vessel group-   20 _(i), 120 _(i) (i=1, . . . , 12) Exclusive regions-   211 _(i) (i=1, . . . , 12) Dispensing tips-   231 _(i), 236 _(i) (i=1, . . . , 12) PCR tubes (reaction vessels)-   30, 300, 130, 1300 Connecting end arranging body-   31 _(i), 131 _(i) (i=1, . . . , 12) Linking portions-   32, 320, 132 Light guide stage-   33 _(i) Optical fibers (example of light guide portions)-   40, 400, 140 Measurement device-   40 _(j), 400 _(j), 1400 _(j) (j=1, . . . , 6) Specific wavelength    measurement devices-   44 Measuring end-   48 _(j) Modulator-   49 _(j) Tuner-   50, 500, 150 Nozzle head-   52 Transfer mechanism portion-   53 Suction-discharge mechanism-   59 Tip detaching mechanism-   61, 161 Measurement control portion-   70 Nozzle arranging portion-   71 _(i) (i=1, . . . , 12) Nozzle

The invention claimed is:
 1. A light measurement apparatus for areaction vessel comprising: two or more linking portions to which isprovided front ends of one or two or more light guide portions that aredirectly or indirectly linkable with two or more reaction vessels andoptically connect with the interior of the reaction vessel to which itis linked; a connecting end arranging body that has an arranging surfacethat arranges and supports along a predetermined path, two or moreconnecting ends, to which is provided back ends of the light guideportions, in which the front ends thereof are provided to the linkingportions, the connecting ends are provided corresponding to therespective linking portions; a measurement device that has one or two ormore measuring ends that are successively optically connectable with therespective two or more connecting ends along the predetermined path onthe arranging surface, and in which light based on an optical statewithin the reaction vessels is receivable by means of opticalconnections between the two or more connecting ends and the measuringends, the measuring ends are provided approaching or making contact withthe arranging surface; and a light guide switching mechanism thatrelatively moves the respective two or more connecting ends arranged onthe connecting end arranging body and the respective measuring endsalong the predetermined path on the arranging surface such that they aresuccessively optically connected, in a state that the two or morelinking portions are held to be linked to the respective two or morereaction vessels, wherein the measurement device comprises a pluralityof types of measurement devices each including a photoelectric element,and provided with respect to the respective measurement devices, and atthe very least between adjacent measurement devices, are a modulatorthat modulates the intensity of the light to be received by thephotoelectric elements provided in the interior of the respectivemeasurement devices at mutually different predetermined frequencies, anda tuner that, with regard to the light received by the respectivemeasurement devices, performs tuning to the predetermined frequencies,and obtains the corresponding intensity of the received light, such thatoptical crosstalk mainly due to the entry of excitation light fromadjacent measurement devices is prevented.
 2. A light measurementapparatus for a reaction vessel according to claim 1, wherein at least aportion of the light guide portion has flexibility.
 3. A lightmeasurement apparatus for a reaction vessel according to claim 1,wherein the measurement device comprises a plurality of types ofspecific wavelength measurement devices that are able to receive therespective light of specific wavelengths or specific wavelength bands,the specific wavelength measurement devices having at least onemeasuring end that is successively optically connectable with theconnecting ends along the predetermined path, and the respectivepredetermined frequencies of the modulators corresponding to thespecific wavelength measurement devices being mutually different, atleast between adjacent specific wavelength measurement devices, and in acase where the light based on the optical state is a fluorescent light,the specific wavelength measurement devices are provided with anirradiation source that irradiates a predetermined excitation light thatexcites the corresponding fluorescent light of a specific wavelength ora specific wavelength band, and a light receiving portion that is ableto receive the fluorescent light of the specific wavelength or thespecific wavelength band, and the modulators that correspond to therespective specific wavelength measurement devices are provided with anexcitation light modulation portion that modulates the predeterminedexcitation lights at the predetermined frequency, and the tuners areprovided with a bandpass filter circuit or a lock-in amplifier that,with respect to the light received by the light receiving portion,extracts the intensity data of the light having the predeterminedfrequencies.
 4. A light measurement apparatus according to claim 1, thathas two or more linking portions provided on a light guide stage, and astage transfer mechanism that relatively moves the light guide stagewith respect to the reaction vessels such that the linking portion issimultaneously directly or indirectly linked with the two or morereaction vessels.
 5. A light measurement apparatus for a reaction vesselaccording to claim 1, provided with a temperature controller having atemperature source that is provided making contact with or approachingthe lower side wall sections of the reaction vessels, and a heatingportion provided such that it is able to make contact with or approachthe upper side wall sections of the reaction vessels positioned furtheron an upper side than the lower side wall sections of the reactionvessels, and that has a heat source that is able to heat the upper sidewall sections.
 6. A light measurement apparatus for a reaction vesselaccording to claim 1, wherein a linking portion, which is directly orindirectly linkable with the reaction vessels and is provided with anoptical element in the interior, is made from a material that is usableto a temperature higher than the dew point of water vapor, and has athermal conductivity smaller than 1.0 W/(m·K).
 7. A light measurementapparatus for a reaction vessel according to claim 1, wherein front endsof a light guide portion bundle which comprises a plurality of lightguide portions, are provided to the respective linking portions, backends of a light guide portion bundle of a portion of the light guideportion bundle are provided to first connecting ends of the connectingend arranging body, a portion or all of the remainder of the light guideportion bundle is provided to second connecting ends of the connectingend arranging body, the predetermined path comprises a first path and asecond path, and the connecting end arranging body has a plurality ofpairs of first connecting ends and second connecting ends that areoptically connected with the linking portion, and a single pair of athird connecting end and a fourth connecting end that is opticallymutually connected by means of the light guide portion, and the firstmeasuring end provided to the measurement device is provided relativelymovable along a first path, which comprises a plurality of the firstconnecting ends and a single third connecting end, and the secondmeasuring end provided to the measurement device is provided relativelymovable along a second path, which comprises a plurality of the secondconnecting ends and a single fourth connecting end, and the firstmeasuring end is optically connect with the irradiation source of themeasurement device, and the second measuring end is connected with thelight receiving portion of the measurement device, and for each pair,the first measuring end is successively connectable with the pluralityof first connecting ends and the single third connecting end, and thesecond measuring end is successively connectable with the plurality ofsecond connecting ends and the single fourth connecting end.
 8. A lightmeasurement apparatus for a reaction vessel comprising: a nozzle headprovided with a suction-discharge mechanism that performs suction anddischarge of gases, and one or two or more nozzles that detachably mountdispensing tips in which the suction and the discharge of liquids ispossible by means of the suction-discharge mechanism; a vessel grouphaving at the very least one or two or more liquid housing parts thathouse reaction solutions used for various reactions, a liquid housingpart that houses a magnetic particle suspension in which magneticparticles that are able to capture a target compound are suspended, aliquid housing part that houses a sample, one or two or more liquidhousing parts that house a solution for separating and extracting of thetarget compound, and two or more reaction vessels; a nozzle headtransfer mechanism that makes an interval between the nozzle head andthe vessel group relatively movable; a magnetic force part that is ableto adsorb the magnetic particles on an inner wall of dispensing tipsmounted on the nozzles; a light guide stage provided to the nozzle headand having two or more linking portions to which ends of one or two ormore light guide portions, which have a flexibility, that are directlyor indirectly linkable with the respective reaction vessels andoptically connect with the interior of the linked reaction vessels, areprovided; a connecting end arranging body having an arranging surfacethat arranges and supports along a predetermined path two or moreconnecting ends, to which back ends of the light guide portions, inwhich front ends thereof are provided to the linking portions, areprovided, the connecting ends are provided corresponding to therespective linking portions; a measurement device having one or two ormore measuring ends that are successively optically connectable with therespective two or more connecting ends along the predetermined path onthe arranging surface, that is able to receive light based on an opticalstate within the reaction vessels by means of optical connectionsbetween the two or more connecting ends and the measuring ends, themeasuring ends are provided approaching or making contact with thearranging surface; and a light guide switching mechanism provided alongthe predetermined path of the connecting end arranging body thatrelatively moves the respective two or more connecting ends and therespective measuring ends along the predetermined path on the arrangingsurface such that they become successively optically connected, in astate that the two or more linking portions are held to be linked to therespective two or more reaction vessels, wherein the measurement devicecomprises a plurality of types of measurement devices each including aphotoelectric element, and the apparatus is configured to at the veryleast between adjacent measurement devices, modulate the intensity ofthe light to be received by the photoelectric elements provided in theinterior of the respective measurement devices at mutually differentpredetermined frequencies, tune the light received by the measurementdevices to the predetermined frequencies, and obtain the correspondingintensity of the received light, such that optical crosstalk mainly dueto the entry of excitation light from adjacent measurement devices isprevented.
 9. A light measurement method for a reaction vesselcomprising: moving two or more linking portions provided with an end ofone or two or more light guide portions, with respect to two or morereaction vessels; simultaneously directly or indirectly linking thereaction vessels and the linking portions and optically connecting theinterior of the linked reaction vessels and the light guide portions;performing temperature control within the reaction vessels; guidinglight from the reaction vessels to a connecting end arranging bodyhaving an arranging surface that arranges and supports along apredetermined path two or more connecting ends which are directly orindirectly optically linked to two or more reaction vessels, and towhich back ends of the light guide portions, in which front ends thereofare provided to the linking portions, are provided, the connecting endsare provided corresponding to the respective linking portions; andsuccessively optically connecting along the predetermined path the oneor two or more measuring ends provided to a measurement device, whichare provided approaching or in contact with the arranging surface, andthe respective two or more connecting ends, in a state that the two ormore linking portions are held to be linked to the respective two ormore reaction vessels, by relatively moving the connecting end arrangingbody along the predetermined path on the arranging surface, to therebymake a plurality of types of measurement devices receive light based onan optical state within the reaction vessels, wherein the measurementdevice comprises a plurality of types of measurement devices eachincluding a photoelectric element, and the method comprises at the veryleast between adjacent measurement devices, modulating the intensity ofthe light to be received by the photoelectric elements provided in theinterior of the respective measurement devices at mutually differentpredetermined frequencies, tuning the light received by the measurementdevices to the predetermined frequencies, and obtaining thecorresponding intensity of the received light, such that opticalcrosstalk mainly due to the entry of excitation light from adjacentmeasurement devices is prevented.
 10. A light measurement method for areaction vessel according to claim 9, wherein an optical connectionbetween the reaction vessel and the connecting ends is directly orindirectly linkable with two or more reaction vessels, and back ends ofone or two or more light guide portions that optically connect with theinterior of the linked reaction vessels, are provided on the connectingends, and the optical connection is performed by relatively moving alight guide stage on which is provided the two or more linking portionsprovided with front ends of the light guide portions, with respect tothe reaction vessel.
 11. A light measurement method for a reactionvessel according to claim 9, wherein the front ends of a light guideportion bundle comprising a plurality of light guide portions areprovided to the respective linking portions, the light guide portionbundle is such that the rear ends of a portion of the light guideportion bundle are provided to first connecting ends of the connectingend arranging body, a portion or all of the remaining rear ends of thelight guide portion bundle are provided to second ends of the connectingend arranging body, the predetermined path comprises a first path and asecond path, the connecting end arranging body has a plurality of pairsof first connecting ends and second connecting ends that are opticallyconnected with the linking portion, and a single pair of a thirdconnecting end and a fourth connecting end that is mutually opticallyconnected by means of the light guide portion, and at the time the firstmeasuring end provided to the measurement device relatively moves alongthe first path, which comprises a plurality of the first connecting endsand a single third connecting end, and the second measuring end providedto the measurement device relatively moves along the second path, whichcomprises the plurality of second connecting ends and a single fourthconnecting end, the first measuring ends optically connect with theirradiation source of the measurement device, and the second measuringends connect with the light receiving portion of the measurement device,and for each pair, the first measuring end successively connects withthe plurality of first connecting ends and the single third connectingend, and the second measuring end successively connects with theplurality of second connecting ends and the single fourth connectingend.
 12. A light measurement method for a reaction vessel according toclaim 9, comprising: directly or indirectly linking an aperture of thereaction vessel and the linking portion; and at the time of performingtemperature control within the reaction vessels, according totemperature control by a temperature controller, which has a temperaturesource provided making contact with or approaching lower side wallsections of the reaction vessels, heating upper side wall sections ofthe reaction vessels, which are positioned further on an upper side thanthe lower side wall sections, by means of a heat source of a heatingportion, which is provided making contact with or approaching the upperside wall sections, and thereby preventing direct or indirectcondensation of the linking portions.
 13. A light measurement method fora reaction vessel comprising: detachably mounting dispensing tips onrespective nozzles, which are provided to a nozzle head and perform thesuction and the discharge of gases; separating a target compound byusing a magnetic force part, a nozzle head transfer mechanism thatrelatively moves an interval between the nozzle head and a vessel group,a magnetic particle suspension housed within a vessel group, in whichmagnetic particles that are able to capture a target compound aresuspended, a sample, and a solution for separating and extracting of atarget compound; introducing the separated target compound and areaction solution used for reactions to a plurality of reaction vesselsprovided to a vessel group; moving a light guide stage, which has two ormore linking portions that are provided to the nozzle head and in whichfront ends of one or two or more light guide portions are also provided,with respect to the reaction vessels by means of, at the very least, thenozzle head transfer mechanism; directly or indirectly simultaneouslylinking the respective reaction vessels and the linking portions, andoptically connecting the interior of the reaction vessels and the lightguide portions that are linked; performing temperature control withinthe reaction vessels; guiding light from the reaction vessels to aconnecting end arranging body that arranges and supports along apredetermined path two or more connecting ends, to which back ends ofthe light guide portions, in which front ends thereof are provided tothe linking portions, are provided, the connecting ends are providedcorresponding to the respective linking portions; and successivelyoptically connecting one or two or more measuring ends that are providedto a measurement device, and provided approaching or making contact withan arranging surface, and the respective two or more connecting ends, ina state that the two or more linking portions are held to be linked tothe respective two or more reaction vessels, along the predeterminedpath by moving the connecting end arranging body, to thereby make themeasurement device receive the light based on an optical state withinthe reaction vessels, wherein the measurement device comprises aplurality of types of measurement devices each including a photoelectricelement, and the method comprises, at the very least between adjacentmeasurement devices, modulating the intensity of the light to bereceived by the photoelectric elements provided in the interior of therespective measurement devices at mutually different predeterminedfrequencies, tuning the light received by the respective measurementdevices to the predetermined frequencies, and obtaining thecorresponding intensity of the received light, such that opticalcrosstalk mainly due to the entry of excitation light from adjacentmeasurement devices is prevented.
 14. A light measurement method for areaction vessel according to claim 13, wherein the measurement devicecomprises a plurality of types of measurement devices, and the methodcomprises at the time of guiding light from the reaction vessels to aconnecting end arranging body, at the very least between adjacentmeasurement devices, modulating the intensity of the light to bereceived by the measurement devices at mutually different predeterminedfrequencies, and at the time the measurement device receives the lightbased on an optical state within the reaction vessels, tuning the lightreceived by the measurement devices to the predetermined frequencies,and obtaining the corresponding intensity of the received light.